Proteins comprising cd3 antigen binding domains and uses thereof

ABSTRACT

The disclosure provides antigen binding domains that bind cluster of differentiation 3 (CD3) protein, comprising the antigen binding domains that bind CD3ε, polynucleotides encoding them, vectors, host cells, methods of making and using them.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application Ser.No. 63/165,184, filed 24 Mar. 2021. The entire contents of theaforementioned application are incorporated herein by reference in itsentirety.

REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

This application contains a sequence listing, which is submittedelectronically via EFS-Web as an ASCII formatted sequence listing with afile name “JBI6516USNP1_SL.txt”, creation date of Mar. 17, 2022 andhaving a size of 1,282 KB. The sequence listing submitted via EFS-Web ispart of the specification and is herein incorporated by reference in itsentirety.

TECHNICAL FIELD

The disclosure provides antigen binding domains that bind cluster ofdifferentiation 3 (CD3) protein comprising the antigen binding domainsthat bind CD3, polynucleotides encoding them, vectors, host cells,methods of making and using them.

BACKGROUND

Bispecific antibodies and antibody fragments have been explored as ameans to recruit cytolytic T cells to kill tumor cells. However, theclinical use of many T cell-recruiting bispecific antibodies has beenlimited by challenges including unfavorable toxicity, potentialimmunogenicity, and manufacturing issues. There thus exists aconsiderable need for improved bispecific antibodies that recruitcytolytic T cells to kill tumor cells that include, for example, reducedtoxicity and favorable manufacturing profiles.

The human CD3 T cell antigen receptor protein complex is composed of sixdistinct chains: a CD3γ chain (SwissProt P09693), a CD3δ chain(SwissProt P04234), two CD3ε chains (SwissProt P07766), and one CD3ζchain homodimer (SwissProt P20963) (εγ:εδ:ζζ), which is associated withthe T cell receptor α and β chain. This complex plays an important rolein coupling antigen recognition to several intracellularsignal-transduction pathways. The CD3 complex mediates signaltransduction, resulting in T cell activation and proliferation. CD3 isrequired for immune response.

Redirection of cytotoxic T cells to kill tumor cells has become animportant therapeutic mechanism for numerous oncologic indications(Labrijn, A. F., Janmaat, M. L., Reichert, J. M. & Parren, P. Bispecificantibodies: a mechanistic review of the pipeline. Nat Rev Drug Discov18, 585-608, doi:10.1038/s41573-019-0028-1 (2019)). T cell activationfollows a two-signal hypothesis, in which the first signal is suppliedby engagement of the T cell receptor (TCR) complex with its cognatepeptide MHC complex on an antigen presenting cell (APC), and the secondsignal may be either co-stimulatory or co-inhibitory (Chen, L. & Flies,D. B. Molecular mechanisms of T cell co-stimulation and co-inhibition.Nat Rev Immunol 13, 227-242, doi:10.1038/nri3405 (2013)). Tumors oftenfail to present sufficient non-self antigens to induce a T cell-basedimmune response, and T cell-engaging BsAbs (bsTCE) can overcome thischallenge by inducing T cell activation in the absence of TCR-pMHCinteraction. T cell receptor signaling occurs through the ITAM motifs inthe cytoplasmic region of the CD3 subunits of the TCR (Chen, D. S. &Mellman, I. Oncology meets immunology: the cancer-immunity cycle.Immunity 39, 1-10, doi:10.1016/j.immuni.2013.07.012 (2013)). Inparticular, the CD3ε subunit is present in two copies per TCR complexand represents an attractive antigen for T cell engagement. Indeed,numerous bsTCE that target CD3ε have shown clinical anti-tumor efficacywhere mAbs have failed, and significant pharmaceutical developmentefforts are ongoing for several tumor targets (Labrijn, A. F. et al.,2019). Three major challenges for clinical development of bsTCE are 1)the potential for rapid and severe toxicity associated with cytokinerelease via systemic or off-tumor T cell activation, 2) practicalchallenges of formulation and dosing for bsTCE with high potency andsharp therapeutic indices, and 3) the potential for reactivation-inducedT cell death, wherein tumor-infiltrating T cells (TILS) undergoapoptosis in response to over-activation by bsTCE (Wu, Z. & Cheung, N.V. T cell engaging bispecific antibody (T-BsAb): From technology totherapeutics. Pharmacol Ther 182, 161-175,doi:10.1016/j.pharmthera.2017.08.005 (2018)).

Together, these observations suggest that there is a need in the art fornovel CD3 specific binding proteins that are more advantageous and canbe used to treat cancers.

SUMMARY

The disclosure satisfies this need, for example, by providing novel CD3εspecific binding proteins that possess high affinity for the tumorantigen and weak affinity for the T cell. The proteins comprising anantigen binding domain that binds CD3ε of the disclosure were generatedto have high thermostability, reduced deamidation risk, and humanized todecrease immunogenicity.

In certain embodiments, the disclosure provides an isolated proteincomprising an antigen binding domain that binds to cluster ofdifferentiation 3ε (CD3ε), wherein the antigen binding domain that bindsCD3ε comprises: an isolated protein comprising an antigen binding domainthat binds to cluster of differentiation 3ε (CD3ε), wherein the antigenbinding domain that binds CD3ε comprises:

a heavy chain complementarity determining region (HCDR) 1, a HCDR2 and aHCDR3 of a heavy chain variable region (VH) of SEQ ID NO: 55 and a lightchain complementarity determining region (LCDR) 1, a LCDR2 and a LCDR3of a light chain variable region (VL) of SEQ ID NO: 59;

the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 55 and theLCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO: 58;

the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 54 and theLCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO: 56; or

the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 48 and theLCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO: 58;

wherein the amino acid in position N106 of SEQ ID NO: 55, 54, or 48 isoptionally substituted with the amino acid selected from the groupconsisting of A, G, S, F, E, T, R, V, I, Y, L, P, Q, and K, wherein theresidue numbering starts from N-terminus of SEQ ID NO: 55, 54, or 48.

In certain embodiments, the disclosure also provides an isolatedprotein, comprising the HCDR1, the HCDR2, the HCDR3, the LCDR1, theLCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 86, 79, 80, and 81,respectively.

In other embodiments, the HCDR1, the HCDR2, the HCDR3, the LCDR1, theLCDR2 and the LCDR3 comprise

SEQ ID NOs: 70, 71, 72, 79, 80, and 81, respectively;

SEQ ID NOs: 70, 71, 87, 79, 80, and 81, respectively; or

SEQ ID NOs: 70, 71, 90, 79, 80, and 81, respectively.

In other embodiments, the antigen binding domain that binds CD3ε is ascFv, a (scFv)2, a Fv, a Fab, a F(ab′)2, a Fd, a dAb or a VHH.

In other embodiments, the antigen binding domain that binds CD3ε is theFab.

In other embodiments, the antigen binding domain that binds CD3ε is thescFv.

In other embodiments, the scFv comprises, from the N- to C-terminus, aVH, a first linker (L1) and a VL (VH-L1-VL) or the VL, the L1 and the VH(VL-L1-VH).

In other embodiments, the L1 comprises

about 5-50 amino acids;

about 5-40 amino acids;

about 10-30 amino acids; or about 10-20 amino acids.

In other embodiments, the L1 comprises an amino acid sequence of SEQ IDNOs: 3-36.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 3.

In other embodiments, the antigen binding domain that binds CD3εcomprises the VH of SEQ ID NOs: 55, 54, or 48 and the VL of SEQ ID NOs:59, 58 or 56.

In other embodiments, the antigen binding domain that binds CD3εcomprises:

the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59;

the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58;

the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 56;

the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 58

the VH of SEQ ID NO: 88 and the VL of SEQ ID NO: 58; or

the VH of SEQ ID NO: 242 and the VL of SEQ ID NO: 58.

In other embodiments, the antigen binding domain that binds CD3εcomprises an amino acid sequence selected from the group consisting ofSEQ ID NOs: 96-126.

In other embodiments, the isolated protein is a multispecific protein.

In other embodiments, the multispecific protein is a bispecific protein.

In other embodiments, the multispecific protein is a trispecificprotein.

In other embodiments, the isolated protein further comprises animmunoglobulin (Ig) constant region or a fragment of the Ig constantregion thereof.

In other embodiments, the fragment of the Ig constant region comprises aFc region.

In other embodiments, the fragment of the Ig constant region comprises aCH2 domain.

In other embodiments, the fragment of the Ig constant region comprises aCH3 domain.

In other embodiments, the fragment of the Ig constant region comprises aCH2 domain and a CH3 domain.

In other embodiments, the fragment of the Ig constant region comprisesat least portion of a hinge, a CH2 domain and a CH3 domain.

In other embodiments, the fragment of the Ig constant region comprises ahinge, a CH2 domain and a CH3 domain.

In other embodiments, the antigen binding domain that binds CD3ε isconjugated to the N-terminus of the Ig constant region or the fragmentof the Ig constant region.

In other embodiments, the antigen binding domain that binds CD3ε isconjugated to the C-terminus of the Ig constant region or the fragmentof the Ig constant region.

In other embodiments, the antigen binding domain that binds CD3ε isconjugated to the Ig constant region or the fragment of the Ig constantregion via a second linker (L2).

In other embodiments, the L2 comprises the amino acid sequence selectedfrom the group consisting of SEQ ID NOs: 3-36.

In other embodiments, the multispecific protein comprises an antigenbinding domain that binds an antigen other than CD3ε.

In other embodiments, the cell antigen is a tumor associated antigen.

In other embodiments, the Ig constant region or the fragment of the Igconstant region is an IgG1, an IgG2, an IgG3 or an IgG4 isotype.

In other embodiments, the Ig constant region or the fragment of the Igconstant region comprises at least one mutation that results in reducedbinding of the protein to a Fcγ receptor (FcγR).

In other embodiments, the at least one mutation that results in reducedbinding of the protein to the FcγR is selected from the group consistingof F234A/L235A, L234A/L235A, L234A/L235A/D265S,V234A/G237A/P238S/H268A/V309L/A330S/P331S, F234A/L235A,S228P/F234A/L235A, N297A, V234A/G237A,K214T/E233P/L234V/L235A/G236-deleted/A327G/P331A/D365E/L358M,H268Q/V309L/A330S/P331S, S267E/L328F, L234F/L235E/D265A,L234A/L235A/G237A/P238S/H268A/A330S/P331S, S228P/F234A/L235A/G237A/P238Sand S228P/F234A/L235A/G236-deleted/G237A/P238S, wherein residuenumbering is according to the EU index.

In other embodiments, the FcγR is FcγRI, FcγRIIA, FcγRIIB or FcγRIII, orany combination thereof.

In other embodiments, the protein comprises at least one mutation in aCH3 domain of the Ig constant region.

In other embodiments, the at least one mutation in the CH3 domain of theIg constant region is selected from the group consisting of T350V,L351Y, F405A, Y407V, T366Y, T366W, T366L, T366L, F405W, T394W, K392L,T394S, T394W, Y407T, Y407A, T366S/L368A/Y407V, L351Y/F405A/Y407V,T366I/K392M/T394W, F405A/Y407V, T366L/K392M/T394W, T366L/K392L/T394W,L351Y/Y407A, L351Y/Y407V, T366A/K409F, T366V/K409F, T366A/K409F,T350V/L351Y/F405A/Y407V and T350V/T366L/K392L/T394W, wherein residuenumbering is according to the EU index.

The disclosure also provides a pharmaceutical composition comprising theisolated protein and a pharmaceutically acceptable carrier.

The disclosure also provides a polynucleotide encoding the isolatedprotein.

The disclosure also provides a vector comprising the polynucleotide.

The disclosure also provides a host cell comprising the vector.

The disclosure also provides a method of producing the isolated protein,comprising culturing the host cell in conditions that the protein isexpressed, and recovering the protein produced by the host cell.

The disclosure also provides a method of treating a cancer in a subject,comprising administering a therapeutically effective amount of theisolated protein to the subject in need thereof to treat the cancer.

The disclosure also provides an anti-idiotypic antibody binding to theisolated protein.

The disclosure also provides an isolated protein of any one of claims1-35 comprising an amino acid sequence selected from the groupconsisting of SEQ ID NOs: 127-157.

The disclosure also provides an isolated protein comprising an antibodyheavy chain of SEQ ID NO: 224 and antibody light chain of SEQ ID NO:226.

BRIEF DESCRIPTIONS OF THE DRAWINGS

The summary, as well as the following detailed description, is furtherunderstood when read in conjunction with the appended drawings. For thepurpose of illustrating the disclosed antibodies and methods, there areshown in the drawings exemplary embodiments of the antibodies andmethods; however, antibodies and methods are not limited to the specificembodiments disclosed. In the drawings:

FIG. 1 shows binding of murine Cris-7 (CD3B1127 and CD31128) and humangermline-grafted Cris-7 variant sequences in scFv format, as determinedby ELISA.

FIGS. 2A and 2B show % of E. coli-expressed scFv clones retaining atleast 75% of binding, determined by ELISA, after the heat shock of 60°C. for the humanized VH pared with murine VL (2A) or for the humanizedVL paired with murine VH (2B); numbers on the X-axis show the residuepositions.

FIG. 3 shows ELISA-based comparison of the binding abilities ofhumanized CD3 specific scFvs containing human-to-mouse back mutations.

FIG. 4 shows binding of CD3B2030 variants, formatted as scFvs, torecombinant CD3 (TRCW5), determined by ELISA; “NtoX” indicates the aminoacid substitutions made in position 106 of the VH (SEQ ID NO: 55),wherein “X” is the amino acid indicated on the Figure.

FIG. 5 shows hydrogen-deuterium exchange rates determined usinghydrogen-deuterium exchange mass spectrometry (HDX-MS) measured for thecomplex of Cris7 (either bi-valent or monovalent) bound to human CD3ε,or the complex of OKT3 bound to human CD3ε (CD3ε:OKT3) (fragment of CD3ε(SEQ ID No: 1) is shown). Underline indicates segments with >30%decrease in deuteration levels in the presence of the antibody, ascompared to CD3ε alone. FIG. 5 discloses SEQ ID NOS 1508, 1509, 1509 and1509, respectively, in order of appearance.

FIG. 6 . Depiction of a exemplary CD79b×CD20×CD3 trispecific antibody.

FIGS. 7A-7D. Binding affinities of selected CD79b×CD3 bsAbs in the HLB-1cell line (FIG. 7A); the OCI-LY10 cell line (FIG. 7B); the Carnaval cellline (FIG. 7C); and the WILL-2 cell line (FIG. 7D). Circles correspondto the 79C3B646 bsAb; triangles correspond to the 79C3B651 bsAb; anddiamonds correspond to the 79C3B601 bsAb.

FIGS. 8A-8D. Binding affinities of selected CD79b×CD20×CD3 trispecificantibodies in the HLB-1 cell line (FIG. 8A); the OCI-LY10 cell line(FIG. 8B); the Carnaval cell line (FIG. 8C); and the WILL-2 cell line(FIG. 8D). Solid circles correspond to the 79C3B646 bsAb control; solidtriangles correspond to the 79C3B651 bsAb control; and solid diamondscorrespond to the 79C3B601 bsAb control. Open triangles correspond totrispecific antibody C923B38; open diamonds correspond to trispecificantibody C923B74; asterisks correspond to trispecific antibody C923B9;and X corresponds to control null trispecific antibody C923B98.

FIGS. 9A-9I. Binding kinetics of selected CD79b×CD3 bsAbs on DLBCL celllines. Binding kinetics of the three selected bsAbs in HBL-1 cells at300 nm (FIG. 9A). Binding kinetics of the three selected bsAbs in HBL-1cells at 60 nm (FIG. 9B). Binding kinetics of the three selected bsAbsin HBL-1 cells at 12 nm (FIG. 9C). Binding kinetics of the threeselected bsAbs in Carnaval cells at 300 nm (FIG. 9D). Binding kineticsof the three selected bsAbs in Carnaval cells at 60 nm (FIG. 9E).Binding kinetics of the three selected bsAbs in Carnaval cells at 12 nm(FIG. 9F). Binding kinetics of the three selected bsAbs in OCI-LY10cells at 300 nm (FIG. 9G). Binding kinetics of the three selected bsAbsin OCI-LY10 cells at 60 nm (FIG. 9H). Binding kinetics of the threeselected bsAbs in OCI-LY10 cells at 12 nm (FIG. 9I). Inverted trianglescorrespond to the 79C3B646 bsAb; diamonds correspond to the 79C3B651bsAb; and squares correspond to the 79C3B601 bsAb.

FIGS. 10A-10I. Binding kinetics of selected CD79b×CD20×CD3 trispecificantibodies on DLBCL cell lines. Binding kinetics of the selectedantibodies in HBL-1 cells at 300 nm (FIG. 10A). Binding kinetics of theselected antibodies in HBL-1 cells at 60 nm (FIG. 10B). Binding kineticsof the selected antibodies in HBL-1 cells at 12 nm (FIG. 10C). Bindingkinetics of the selected antibodies in Carnaval cells at 300 nm (FIG.10D). Binding kinetics of the selected antibodies in Carnaval cells at60 nm (FIG. 10E). Binding kinetics of the selected antibodies inCarnaval cells at 12 nm (FIG. 10F). Binding kinetics of the selectedantibodies in OCI-LY10 cells at 300 nm (FIG. 10G). Binding kinetics ofthe selected antibodies in OCI-LY10 cells at 60 nm (FIG. 10H). Bindingkinetics of the selected antibodies in OCI-LY10 cells at 12 nm (FIG.10I). Inverted triangles correspond to the 79C3B646 bsAb control;diamonds correspond to the 79C3B651 bsAb control; and squares correspondto the 79C3B601 bsAb control. Triangles correspond to trispecificantibody C923B38; circles correspond to trispecific antibody C923B74;squares correspond to trispecific antibody C923B99; and asteriskscorrespond to control null trispecific antibody C923B98.

FIGS. 11A-11D. Primary pan T-cell binding of CD79b×CD20×CD3 trispecificantibodies and CD79b×CD3 bispecific antibodies. Binding kinetics of theselected antibodies in pan T-cell donor line D221837 (FIG. 11A). Bindingkinetics of the selected antibodies in pan T-cell donor line D329312(FIG. 11B). Binding kinetics of the selected antibodies in pan T-celldonor line D329335 (FIG. 11C). Binding kinetics of the selectedantibodies in pan T-cell donor line D160115 (FIG. 11D). Circlescorrespond to the 79C3B651 bsAb; squares correspond to the 79C3B646bsAb; triangles correspond to the trispecific antibody C923B38; invertedtriangles correspond to the trispecific antibody C923B99; diamondscorrespond to the trispecific antibody C923B74.

FIGS. 12A-12B. T cell cytotoxicity of CD79b×CD20×CD3 trispecificantibodies and CD79b×CD3 bispecific antibodies. Cytotoxicity of theselected antibodies in the HEL T-cell line (FIG. 12A). Cytotoxicity ofthe selected antibodies in the K562 T-cell line (FIG. 12B). Shadedcircles correspond to the trispecific antibody C923B74; clear circlescorrespond to the trispecific antibody C923B99; triangles correspond tothe trispecific antibody C923B38; inverted triangles correspond to the79C3B646 bsAb; diamonds correspond to 79C3B651 bsAb; black squarescorrespond to the 79C3B601 bsAb; and white squares correspond to C923B98bsAb.

FIGS. 13A-13C. CD79b×CD20×CD3 trispecific construct mediated B cellcytotoxicity and T cell activation. Cytotoxicity in B cells (FIG. 13A);CD4⁺ T-cells (FIG. 13B) and CD8⁺ T-cells are shown for the leadantibodies.

FIG. 14 shows HDX-MS epitope mapping of PSMA against PS3B1352 (top) andPS3B1353 (bottom). G is glycosylation site. Black box is epitope andgray is probable epitope. White box indicates no/little change indeuteration level in the presence of the antibody. The residues withoutbox indicate the HDX behaviors were not monitored, because there is nopeptide to cover the residues or the residues are the first two residuesof a peptide. The epitopes of PS3B1352 and PS3B1353 are identical. Theepitopes are residue 597-598 (CR), because the segment 597-598 weresignificantly protected upon binding (average differences in deuterationlevels >10%). The epitopes include residues 599 (D), 602-603 (VV) and605 (R), because they are marginally protected upon binding (averagedifferences in deuteration levels 5%-10%) and are likely to show biggerprotections if longer time points were monitored. The epitopes can belarger than these four segments, because the segments around these foursegments, such as 593-594 (LP), 595 (F), 596 (D), 600 (Y), 601 (A), 604(L), 606-607 (KY), 607-609 (YAD), and 610-612, (KIY), did not exchangeat all in the time window employed and can be protected if longer timepoints were monitored. FIG. 14 discloses SEQ ID NO: 1510.

FIG. 15 shows HDX-MS identified epitopes of PSMA overlaid on X-raycrystal structure. Blue: epitope; sky blue: probable epitope; and cyan:potential epitope.

FIG. 16 shows PAN-T cell binding assay. Human PAN-T cells were treatedwith various concentrations of PSMA/CD3 bispecific antibodies andincubated at 37° C. for 30 minutes followed by CD3 cell surfaceexpression analysis by flow cytometry.

FIG. 17 shows the non-linear regression fit of four-parameter functionof PSMA ligand binding of C4-2B human prostate tumor cells.

FIG. 18 shows a target cell binding assay. C4-2B human prostate tumorcells were treated with various concentrations of PSMA/CD3 bispecificantibodies and incubated at 37° C. for 30 minutes followed by PSMA cellsurface expression analysis by flow cytometry.

FIG. 19 shows internalization of PSMA. Human C4-2B prostate tumor cellswere incubated with PSMA/CD3 bispecific antibodies conjugated toIncuCyte® Human Fab-fluor-pH Red Antibody Labeling Dye for 24 hours.

FIGS. 20A-H show bispecific anti-PSMA/anti-T cell redirection antibodiesevaluated in an IncuCyte®-based cytotoxicity assay. Isolated PAN-T cellswere co-incubated with PSMA+C4-2B cells in the presence of bispecificPSMA/T cell redirection antibodies for 120 hours. Shown are data for (A)PS3B1352, (B) PS3B1356, (C) PS3B1353, (D) PS3B1357, (E) PS3B1354, (F)PS3B937, (G) PS3B1355, and (H) PS3B1358.

FIG. 21 shows T cell redirected killing assay. Normal human PBMCs werecombined with C4-2B human prostate tumor cells transduced with IncuCyte®NucLight red nuclear dye and treated with PSMA/CD3 bispecific antibodiesfor 5 days.

FIG. 22 shows cytokine induction by bispecific anti-PSMA/anti-T cellredirection antibodies. Isolated PAN-T cells were co-incubated withPSMA+C4-2B cells in the presence of bispecific anti-PSMA/anti-T cellredirection antibodies for the indicated time points. IFN-gammaconcentration was measured from supernatants collected at the indicatedtime points.

DETAILED DESCRIPTION OF THE INVENTION

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as though fully set forth.

It is to be understood that the terminology used herein is for thepurpose of describing particular embodiments only and is not intended tobe limiting. Unless defined otherwise, all technical and scientificterms used herein have the same meaning as commonly understood by one ofordinary skill in the art to which the invention pertains.

Although any methods and materials similar or equivalent to thosedescribed herein may be used in the practice for testing of the presentinvention, exemplary materials and methods are described herein. Indescribing and claiming the present invention, the following terminologywill be used.

When a list is presented, unless stated otherwise, it is to beunderstood that each individual element of that list, and everycombination of that list, is a separate embodiment. For example, a listof embodiments presented as “A, B, or C” is to be interpreted asincluding the embodiments, “A,” “B,” “C,” “A or B,” “A or C,” “B or C,”or “A, B, or C.”

As used in this specification and the appended claims, the singularforms “a,” “an,” and “the” include plural referents unless the contentclearly dictates otherwise. Thus, for example, reference to “a cell”includes a combination of two or more cells, and the like.

The transitional terms “comprising,” “consisting essentially of,” and“consisting of” are intended to connote their generally acceptedmeanings in the patent vernacular; that is, (i) “comprising,” which issynonymous with “including,” “containing,” or “characterized by,” isinclusive or open-ended and does not exclude additional, unrecitedelements or method steps; (ii) “consisting of” excludes any element,step, or ingredient not specified in the claim; and (iii) “consistingessentially of” limits the scope of a claim to the specified materialsor steps “and those that do not materially affect the basic and novelcharacteristic(s)” of the claimed invention. Embodiments described interms of the phrase “comprising” (or its equivalents) also provide asembodiments those independently described in terms of “consisting of”and “consisting essentially of.”

“About” means within an acceptable error range for the particular valueas determined by one of ordinary skill in the art, which will depend inpart on how the value is measured or determined, i.e., the limitationsof the measurement system. Unless explicitly stated otherwise within theExamples or elsewhere in the Specification in the context of aparticular assay, result or embodiment, “about” means within onestandard deviation per the practice in the art, or a range of up to 5%,whichever is larger.

“Activation” or “stimulation” or “activated” or “stimulated” refers toinduction of a change in the biologic state of a cell resulting inexpression of activation markers, cytokine production, proliferation ormediating cytotoxicity of target cells. Cells may be activated byprimary stimulatory signals. Co-stimulatory signals can amplify themagnitude of the primary signals and suppress cell death followinginitial stimulation resulting in a more durable activation state andthus a higher cytotoxic capacity. A “co-stimulatory signal” refers to asignal, which in combination with a primary signal, such as TCR/CD3ligation, leads to T cell and/or NK cell proliferation and/orupregulation or downregulation of key molecules.

“Alternative scaffold” refers to a single chain protein framework thatcontains a structured core associated with variable domains of highconformational tolerance. The variable domains tolerate variation to beintroduced without compromising scaffold integrity, and hence thevariable domains can be engineered and selected for binding to aspecific antigen.

“Antibody-dependent cellular cytotoxicity”, “antibody-dependentcell-mediated cytotoxicity” or “ADCC” refers to the mechanism ofinducing cell death that depends upon the interaction of antibody-coatedtarget cells with effector cells possessing lytic activity, such asnatural killer cells (NK), monocytes, macrophages and neutrophils via Fcgamma receptors (FcγR) expressed on effector cells.

“Antibody-dependent cellular phagocytosis” or “ADCP” refers to themechanism of elimination of antibody-coated target cells byinternalization by phagocytic cells, such as macrophages or dendriticcells.

“Antigen” refers to any molecule (e.g., protein, peptide,polysaccharide, glycoprotein, glycolipid, nucleic acid, portionsthereof, or combinations thereof) capable of being bound by an antigenbinding domain or a T-cell receptor that is capable of mediating animmune response. Exemplary immune responses include antibody productionand activation of immune cells, such as T cells, B cells or NK cells.Antigens may be expressed by genes, synthetized, or purified frombiological samples such as a tissue sample, a tumor sample, a cell or afluid with other biological components, organisms, subunits ofproteins/antigens, killed or inactivated whole cells or lysates.

“Antigen binding fragment” or “antigen binding domain” refers to aportion of the protein that binds an antigen. Antigen binding fragmentsmay be synthetic, enzymatically obtainable or genetically engineeredpolypeptides and include portions of an immunoglobulin that bind anantigen, such as the VH, the VL, the VH and the VL, Fab, Fab′, F(ab′)₂,Fd and Fv fragments, domain antibodies (dAb) consisting of one VH domainor one VL domain, shark variable IgNAR domains, camelized VH domains,VHH domains, minimal recognition units consisting of the amino acidresidues that mimic the CDRs of an antibody, such as FR3-CDR3-FR4portions, the HCDR1, the HCDR2 and/or the HCDR3 and the LCDR1, the LCDR2and/or the LCDR3, alternative scaffolds that bind an antigen, andmultispecific proteins comprising the antigen binding fragments. Antigenbinding fragments (such as VH and VL) may be linked together via asynthetic linker to form various types of single antibody designs wherethe VH/VL domains may pair intramolecularly, or intermolecularly inthose cases when the VH and VL domains are expressed by separate singlechains, to form a monovalent antigen binding domain, such as singlechain Fv (scFv) or diabody. Antigen binding fragments may also beconjugated to other antibodies, proteins, antigen binding fragments oralternative scaffolds which may be monospecific or multispecific toengineer bispecific and multispecific proteins.

“Antibodies” is meant in a broad sense and includes immunoglobulinmolecules including monoclonal antibodies including murine, human,humanized and chimeric monoclonal antibodies, antigen binding fragments,multispecific antibodies, such as bispecific, trispecific, tetraspecificetc., dimeric, tetrameric or multimeric antibodies, single chainantibodies, domain antibodies and any other modified configuration ofthe immunoglobulin molecule that comprises an antigen binding site ofthe required specificity. “Full length antibodies” are comprised of twoheavy chains (HC) and two light chains (LC) inter-connected by disulfidebonds as well as multimers thereof (e.g. IgM). Each heavy chain iscomprised of a heavy chain variable region (VH) and a heavy chainconstant region (comprised of domains CH1, hinge, CH2 and CH3). Eachlight chain is comprised of a light chain variable region (VL) and alight chain constant region (CL). The VH and the VL regions may befurther subdivided into regions of hypervariability, termedcomplementarity determining regions (CDR), interspersed with frameworkregions (FR). Each VH and VL is composed of three CDRs and four FRsegments, arranged from amino-to-carboxy-terminus in the followingorder: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. Immunoglobulins may beassigned to five major classes, IgA, IgD, IgE, IgG and IgM, depending onthe heavy chain constant domain amino acid sequence. IgA and IgG arefurther sub-classified as the isotypes IgA1, IgA2, IgG1, IgG2, IgG3 andIgG4. Antibody light chains of any vertebrate species may be assigned toone of two clearly distinct types, namely kappa (κ) and lambda (λ),based on the amino acid sequences of their constant domains.

“Bispecific” refers to a molecule (such as a protein or an antibody)that specifically binds two distinct antigens or two distinct epitopeswithin the same antigen. The bispecific molecule may havecross-reactivity to other related antigens, for example to the sameantigen from other species (homologs), such as human or monkey, forexample Macaca cynomolgus (cynomolgus, cyno) or Pan troglodytes, or maybind an epitope that is shared between two or more distinct antigens.

“Bispecific anti-PSMA/anti-CD3 antibody”, “PSMA/CD3 antibody”, “PSMAxCD3antibody,” “anti-PSMA/anti-CD3 protein,” and the like refer to anantibody that binds PSMA and CD3 and that comprises at least one bindingdomain specifically binding PSMA and at least one binding domainspecifically binding CD3. The domains specifically binding PSMA and CD3are typically VH/VL pairs. The bispecific anti-PSMAxCD3 antibody may bemonovalent in terms of its binding to either PSMA or CD3.

“Bispecific anti-CD79b/anti-CD3 antibody”, “anti-CD79b×CD3”, “CD79b/CD3antibody”, “CD79b×CD3 antibody,” “anti-CD79b/anti-CD3 protein,” and thelike refer to an antibody that binds CD79b and CD3 and that comprises atleast one binding domain specifically binding CD79b and at least onebinding domain specifically binding CD3. The domains specificallybinding CD79b and CD3 are typically V_(H)/V_(L) pairs. The bispecificanti CD79b×CD3 antibody may be monovalent in terms of its binding toeither CD79b or CD3.

“Cancer” refers to a broad group of various diseases characterized bythe uncontrolled growth of abnormal cells in the body. Unregulated celldivision and growth results in the formation of malignant tumors thatinvade neighboring tissues and may also metastasize to distant parts ofthe body through the lymphatic system or bloodstream. A “cancer” or“cancer tissue” can include a tumor.

“Cluster of Differentiation 3 ε” or “CD3ε” refers to a known proteinwhich is also called “T-cell surface glycoprotein CD3 epsilon chain”, or“T3E”. CD3ε, together with CD3-gamma, -delta and -zeta, and the T-cellreceptor alpha/beta and gamma/delta heterodimers, forms the T-cellreceptor-CD3 complex. This complex plays an important role in couplingantigen recognition to several intracellular signal-transductionpathways. The CD3 complex mediates signal transduction, resulting in Tcell activation and proliferation. CD3 is required for the immuneresponse. The amino acid sequence of a full length CD3ε is shown in SEQID NO: 1. The amino acid sequence of the extracellular domain (ECD) ofCD3ε is shown in SEQ ID NO: 2. Throughout the specification,“CD3ε-specific” or “specifically binds CD3ε” or “anti-CD3ε antibody”refers to antibodies that bind specifically to the CD3ε polypeptide (SEQID NO: 1), including antibodies that bind specifically to the CD3εextracellular domain (ECD) (SEQ ID NO: 2).

(Human CD3 epsilon) SEQ ID NO: 1MQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQR DLYSGLNQRRI(Human CD3 epsilon extracellular domain) SEQ ID NO: 2DGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCE NCMEMD

“Cluster of Differentiation CD79B protein” or “CD79b” refers to a B-cellantigen receptor (BCR) signaling component Igβ. The amino acid sequencesof the various isoforms are retrievable from GenBank accession numbersAAH32651.1, EAW94232.1, AAH02975.2, NP_000617.1, and NP_001035022.1. Theamino acid sequence of the full length CD79b sequence is shown below(SEQ ID NO: 241). The sequence includes the extracellular domain(residues 29-159) and the cytoplasmic domain (residues 181-229).

(SEQ ID NO: 241) MARLALSPVPSHWMVALLLLLSAEPVPAARSEDRYRNPKGSACSRIWQSPRFIARKRGFTVKMHCYMNSASGNVSWLWKQEMDENPQQLKLEKGRMEESQNESLATLTIQGIRFEDNGIYFCQQKCNNTSEVYQGCGTELRVMGFSTLAQLKQRNTLKDGIIMIQTLLIILFIIVPIFLLLDKDDSKAGMEEDHTYEGLDIDQTATYEDIVTLRTGEVKWSVGEHPGQE.

“Complement-dependent cytotoxicity” or “CDC”, refers to the mechanism ofinducing cell death in which the Fc effector domain of a target-boundprotein binds and activates complement component C1q which in turnactivates the complement cascade leading to target cell death.Activation of complement may also result in deposition of complementcomponents on the target cell surface that facilitate CDC by bindingcomplement receptors (e.g., CR3) on leukocytes.

“Complementarity determining regions” (CDR) are antibody regions thatbind an antigen. There are three CDRs in the VH (HCDR1, HCDR2, HCDR3)and three CDRs in the VL (LCDR1, LCDR2, LCDR3). CDRs may be definedusing various delineations such as Kabat (Wu et al. (1970) J Exp Med132: 211-50; Kabat et al., Sequences of Proteins of ImmunologicalInterest, 5th Ed. Public Health Service, National Institutes of Health,Bethesda, Md., 1991), Chothia (Chothia et al. (1987) J Mol Biol 196:901-17), IMGT (Lefranc et al. (2003) Dev Comp Immunol 27: 55-77) and AbM(Martin and Thornton J Bmol Biol 263: 800-15, 1996). The correspondencebetween the various delineations and variable region numbering isdescribed (see e.g. Lefranc et al. (2003) Dev Comp Immunol 27: 55-77;Honegger and Pluckthun, J Mol Biol (2001) 309:657-70; InternationalImMunoGeneTics (IMGT) database; Web resources (for example, can beretrieved from the Internet <URL: http://www.imgt.org>)). Availableprograms such as abYsis by UCL Business PLC may be used to delineateCDRs. The term “CDR”, “HCDR1”, “HCDR2”, “HCDR3”, “LCDR1”, “LCDR2” and“LCDR3” as used herein includes CDRs defined by any of the methodsdescribed supra, Kabat, Chothia, IMGT or AbM, unless otherwiseexplicitly stated in the specification.

“Decrease,” “lower,” “lessen,” “reduce,” or “abate” refers generally tothe ability of a test molecule to mediate a reduced response (i.e.,downstream effect) when compared to the response mediated by a controlor a vehicle. Exemplary responses are T cell expansion, T cellactivation or T-cell mediated tumor cell killing or binding of a proteinto its antigen or receptor, enhanced binding to a Fcγ or enhanced Fceffector functions such as enhanced ADCC, CDC and/or ADCP. Decrease maybe a statistically significant difference in the measured responsebetween the test molecule and the control (or the vehicle), or adecrease in the measured response, such as a decrease of about 1.1, 1.2,1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20 or 30 fold or more, such as 500,600, 700, 800, 900 or 1000 fold or more (including all integers anddecimal points in between and above 1, e.g., 1.5, 1.6, 1.7. 1.8, etc.).

“Differentiation” refers to a method of decreasing the potency orproliferation of a cell or moving the cell to a more developmentallyrestricted state.

“Encode” or “encoding” refers to the inherent property of specificsequences of nucleotides in a polynucleotide, such as a gene, a cDNA, oran mRNA, to serve as templates for synthesis of other polymers andmacromolecules in biological processes having either a defined sequenceof nucleotides (e.g., rRNA, tRNA and mRNA) or a defined sequence ofamino acids and the biological properties resulting therefrom. Thus, agene, cDNA, or RNA, encodes a protein if transcription and translationof mRNA corresponding to that gene produces the protein in a cell orother biological system. Both the coding strand, the nucleotide sequenceof which is identical to the mRNA sequence, and the non-coding strand,used as the template for transcription of a gene or cDNA, can bereferred to as encoding the protein or other product of that gene orcDNA.

“Enhance,” “promote,” “increase,” “expand” or “improve” refers generallyto the ability of a test molecule to mediate a greater response (i.e.,downstream effect) when compared to the response mediated by a controlor a vehicle. Exemplary responses are T cell expansion, T cellactivation or T-cell mediated tumor cell killing or binding of a proteinto its antigen or receptor, enhanced binding to a Fcγ or enhanced Fceffector functions such as enhanced ADCC, CDC and/or ADCP Enhance may bea statistically significant difference in the measured response betweenthe test molecule and control (or vehicle), or an increase in themeasured response, such as an increase of about 1.1, 1.2, 1.5, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20 or 30 fold or more, such as 500, 600, 700,800, 900 or 1000 fold or more (including all integers and decimal pointsin between and above 1, e.g., 1.5, 1.6, 1.7. 1.8, etc.).

“Epitope” refers to a portion of an antigen to which an antibody, or theantigen binding portion thereof, specifically binds. Epitopes typicallyconsist of chemically active (such as polar, non-polar or hydrophobic)surface groupings of moieties such as amino acids or polysaccharide sidechains and may have specific three-dimensional structuralcharacteristics, as well as specific charge characteristics. An epitopemay be composed of contiguous and/or discontiguous amino acids that forma conformational spatial unit. For a discontiguous epitope, amino acidsfrom differing portions of the linear sequence of the antigen come inclose proximity in 3-dimensional space through the folding of theprotein molecule. Antibody “epitope” depends on the methodology used toidentify the epitope.

“Expansion” refers to the outcome of cell division and cell death.

“Express” and “expression” refers the to the well-known transcriptionand translation occurring in cells or in vitro. The expression product,e.g., the protein, is thus expressed by the cell or in vitro and may bean intracellular, extracellular or a transmembrane protein.

“Expression vector” refers to a vector that can be utilized in abiological system or in a reconstituted biological system to direct thetranslation of a polypeptide encoded by a polynucleotide sequencepresent in the expression vector.

“dAb” or “dAb fragment” refers to an antibody fragment composed of a VHdomain (Ward et al., Nature 341:544 546 (1989)).

“Fab” or “Fab fragment” refers to an antibody fragment composed of VH,CH1, VL and CL domains.

“F(ab′)₂” or “F(ab′)₂ fragment” refers to an antibody fragmentcontaining two Fab fragments connected by a disulfide bridge in thehinge region.

“Fd” or “Fd fragment” refers to an antibody fragment composed of VH andCH1 domains.

“Fv” or “Fv fragment” refers to an antibody fragment composed of the VHand the VL domains from a single arm of the antibody.

“Full length antibody” is comprised of two heavy chains (HC) and twolight chains (LC) inter-connected by disulfide bonds as well asmultimers thereof (e.g. IgM). Each heavy chain is comprised of a heavychain variable domain (VH) and a heavy chain constant domain, the heavychain constant domain comprised of subdomains CH1, hinge, CH2 and CH3.Each light chain is comprised of a light chain variable domain (VL) anda light chain constant domain (CL). The VH and the VL may be furthersubdivided into regions of hypervariability, termed complementaritydetermining regions (CDR), interspersed with framework regions (FR).Each VH and VL is composed of three CDRs and four FR segments, arrangedfrom amino-to-carboxy-terminus in the following order: FR1, CDR1, FR2,CDR2, FR3, CDR3 and FR4.

“Genetic modification” refers to the introduction of a “foreign” (i.e.,extrinsic or extracellular) gene, DNA or RNA sequence to a host cell, sothat the host cell will express the introduced gene or sequence toproduce a desired substance, typically a protein or enzyme coded by theintroduced gene or sequence. The introduced gene or sequence may also becalled a “cloned” or “foreign” gene or sequence, may include regulatoryor control sequences operably linked to polynucleotide encoding thechimeric antigen receptor, such as start, stop, promoter, signal,secretion, or other sequences used by a cell's genetic machinery. Thegene or sequence may include nonfunctional sequences or sequences withno known function. A host cell that receives and expresses introducedDNA or RNA has been “genetically engineered.” The DNA or RNA introducedto a host cell can come from any source, including cells of the samegenus or species as the host cell, or from a different genus or species.

“Heterologous” refers to two or more polynucleotides or two or morepolypeptides that are not found in the same relationship to each otherin nature.

“Heterologous polynucleotide” refers to a non-naturally occurringpolynucleotide that encodes two or more neoantigens as described herein.

“Heterologous polypeptide” refers to a non-naturally occurringpolypeptide comprising two or more neoantigen polypeptides as describedherein.

“Host cell” refers to any cell that contains a heterologous nucleicacid. An exemplary heterologous nucleic acid is a vector (e.g., anexpression vector).

“Human antibody” refers to an antibody that is optimized to have minimalimmune response when administered to a human subject. Variable regionsof human antibody are derived from human immunoglobulin sequences. Ifhuman antibody contains a constant region or a portion of the constantregion, the constant region is also derived from human immunoglobulinsequences. Human antibody comprises heavy and light chain variableregions that are “derived from” sequences of human origin if thevariable regions of the human antibody are obtained from a system thatuses human germline immunoglobulin or rearranged immunoglobulin genes.Such exemplary systems are human immunoglobulin gene libraries displayedon phage, and transgenic non-human animals such as mice or rats carryinghuman immunoglobulin loci. “Human antibody” typically contains aminoacid differences when compared to the immunoglobulins expressed inhumans due to differences between the systems used to obtain the humanantibody and human immunoglobulin loci, introduction of somaticmutations or intentional introduction of substitutions into theframeworks or CDRs, or both. Typically, “human antibody” is at leastabout 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98% or 99% identical in amino acid sequence toan amino acid sequence encoded by human germline immunoglobulin orrearranged immunoglobulin genes. In some cases, “human antibody” maycontain consensus framework sequences derived from human frameworksequence analyses, for example as described in Knappik et al., (2000) JMol Biol 296:57-86, or a synthetic HCDR3 incorporated into humanimmunoglobulin gene libraries displayed on phage, for example asdescribed in Shi et al., (2010) J Mol Biol 397:385-96, and in Int.Patent Publ. No. WO2009/085462. Antibodies in which at least one CDR isderived from a non-human species are not included in the definition of“human antibody”.

“Humanized antibody” refers to an antibody in which at least one CDR isderived from non-human species and at least one framework is derivedfrom human immunoglobulin sequences. Humanized antibody may includesubstitutions in the frameworks so that the frameworks may not be exactcopies of expressed human immunoglobulin or human immunoglobulingermline gene sequences.

“In combination with” means that two or more therapeutic agents are beadministered to a subject together in a mixture, concurrently as singleagents or sequentially as single agents in any order.

“Intracellular signaling domain” or “cytoplasmic signaling domain”refers to an intracellular portion of a molecule. It is the functionalportion of the protein which acts by transmitting information within thecell to regulate cellular activity via defined signaling pathways bygenerating second messengers or functioning as effectors by respondingto such messengers. The intracellular signaling domain generates asignal that promotes an immune effector function of the CAR containingcell, e.g., a CAR-T cell.

“Isolated” refers to a homogenous population of molecules (such assynthetic polynucleotides or polypeptides) which have been substantiallyseparated and/or purified away from other components of the system themolecules are produced in, such as a recombinant cell, as well as aprotein that has been subjected to at least one purification orisolation step. “Isolated” refers to a molecule that is substantiallyfree of other cellular material and/or chemicals and encompassesmolecules that are isolated to a higher purity, such as to 80%, 81%,82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or 100% purity.

“Modulate” refers to either enhanced or decreased ability of a testmolecule to mediate an enhanced or a reduced response_(i.e., downstreameffect) when compared to the response mediated by a control or avehicle.

“Monoclonal antibody” refers to an antibody obtained from asubstantially homogenous population of antibody molecules, i.e., theindividual antibodies comprising the population are identical except forpossible well-known alterations such as removal of C-terminal lysinefrom the antibody heavy chain or post-translational modifications suchas amino acid isomerization or deamidation, methionine oxidation orasparagine or glutamine deamidation. Monoclonal antibodies typicallybind one antigenic epitope. A bispecific monoclonal antibody binds twodistinct antigenic epitopes. Monoclonal antibodies may haveheterogeneous glycosylation within the antibody population. Monoclonalantibody may be monospecific or multispecific such as bispecific,monovalent, bivalent or multivalent.

“Multispecific” refers to a molecule, such as an antibody thatspecifically binds two or more distinct antigens or two or more distinctepitopes within the same antigen. Multispecific molecule may havecross-reactivity to other related antigens, for example to the sameantigen from other species (homologs), such as human or monkey, forexample Macaca fascicularis (cynomolgus, cyno) or Pan troglodytes, ormay bind an epitope that is shared between two or more distinctantigens.

“Natural killer cell” and “NK cell” are used interchangeably andsynonymously herein. NK cell refers to a differentiated lymphocyte witha CD16⁺CD56⁺ and/or CD57⁺ TCR: phenotype. NK cells are characterized bytheir ability to bind to and kill cells that fail to express “self”MHC/HLA antigens by the activation of specific cytolytic enzymes, theability to kill tumor cells or other diseased cells that express aligand for NK activating receptors, and the ability to release proteinmolecules called cytokines that stimulate or inhibit the immuneresponse.

“Operatively linked” and similar phrases, when used in reference tonucleic acids or amino acids, refers to the operational linkage ofnucleic acid sequences or amino acid sequence, respectively, placed infunctional relationships with each other. For example, an operativelylinked promoter, enhancer elements, open reading frame, 5′ and 3′ UTR,and terminator sequences result in the accurate production of a nucleicacid molecule (e.g., RNA) and in some instances to the production of apolypeptide (i.e., expression of the open reading frame). Operativelylinked peptide refers to a peptide in which the functional domains ofthe peptide are placed with appropriate distance from each other toimpart the intended function of each domain.

The term “paratope” refers to the area or region of an antibody moleculewhich is involved in binding of an antigen and comprise residues thatinteract with an antigen. A paratope may composed of continuous and/ordiscontinuous amino acids that form a conformational spatial unit. Theparatope for a given antibody can be defined and characterized atdifferent levels of details using a variety of experimental andcomputational methods. The experimental methods includehydrogen/deuterium exchange mass spectrometry (HX-MS). The paratope willbe defined differently depending on the mapping method employed.

“Pharmaceutical combination” refers to a combination of two or moreactive ingredients administered either together or separately.

“Pharmaceutical composition” refers to a composition that results fromcombining an active ingredient and a pharmaceutically acceptablecarrier.

“Pharmaceutically acceptable carrier” or “excipient” refers to aningredient in a pharmaceutical composition, other than the activeingredient, which is nontoxic to a subject. Exemplary pharmaceuticallyacceptable carriers are a buffer, stabilizer or preservative.

“Polynucleotide” or “nucleic acid” refers to a synthetic moleculecomprising a chain of nucleotides covalently linked by a sugar-phosphatebackbone or other equivalent covalent chemistry. cDNA is a typicalexample of a polynucleotide. Polynucleotide may be a DNA or a RNAmolecule.

“Prevent,” “preventing,” “prevention,” or “prophylaxis” of a disease ordisorder means preventing that a disorder occurs in a subject.

“Proliferation” refers to an increase in cell division, either symmetricor asymmetric division of cells.

“Promoter” refers to the minimal sequences required to initiatetranscription. Promoter may also include enhancers or repressor elementswhich enhance or suppress transcription, respectively.

“Protein” or “polypeptide” are used interchangeably herein and refer toa molecule that comprises one or more polypeptides each comprised of atleast two amino acid residues linked by a peptide bond. Protein may be amonomer, or may be protein complex of two or more subunits, the subunitsbeing identical or distinct. Small polypeptides of less than 50 aminoacids may be referred to as “peptides”. Protein may be a heterologousfusion protein, a glycoprotein, or a protein modified bypost-translational modifications such as phosphorylation, acetylation,myristoylation, palmitoylation, glycosylation, oxidation, formylation,amidation, citrullination, polyglutamylation, ADP-ribosylation,pegylation or biotinylation. Protein may be an antibody or may comprisean antigen binding fragment of an antibody. Protein may be recombinantlyexpressed.

“Prostate-specific membrane antigen” or “PSMA” refers to a type IImembrane protein expressed on certain cells. The amino acid sequence ofthe human PSMA is shown in SEQ ID NO: 240. The extracellular domainspans residues 44-750, the transmembrane domain spans residues 20-43 andthe cytoplasmic domain spans residues 1-19 of SEQ ID NO: 240.

SEQ ID NO: 240 MWNLLHETDSAVATARRPRWLCAGALVLAGGFFLLGFLFGWFIKSSNEATNITPKHNMKAFLDELKAENIKKFLYNFTQIPHLAGTEQNFQLAKQIQSQWKEFGLDSVELAHYDVLLSYPNKTHPNYISIINEDGNEIFNTSLFEPPPPGYENVSDIVPPFSAFSPQGMPEGDLVYVNYARTEDFFKLERDMKINCSGKIVIARYGKVFRGNKVKNAQLAGAKGVILYSDPADYFAPGVKSYPDGWNLPGGGVQRGNILNLNGAGDPLTPGYPANEYAYRRGIAEAVGLPSIPVHPIGYYDAQKLLEKMGGSAPPDSSWRGSLKVPYNVGPGFTGNFSTQKVKMHIHSTNEVTRIYNVIGTLRGAVEPDRYVILGGHRDSWVFGGIDPQSGAAVVHEIVRSFGTLKKEGWRPRRTILFASWDAEEFGLLGSTEWAEENSRLLQERGVAYINADSSIEGNYTLRVDCTPLMYSLVHNLTKELKSPDEGFEGKSLYESWTKKSPSPEFSGMPRISKLGSGNDFEVFFQRLGIASGRARYTKNWETNKFSGYPLYHSVYETYELVEKFYDPMFKYHLTVAQVRGGMVFELANSIVLPFDCRDYAVVLRKYADKIYSISMKHPQEMKTYSVSFDSLFSAVKNFTEIASKFSERLQDFDKSNPIVLRMMNDQLMFLERAFIDPLGLPDRPFYRHVIYAPSSHNKYAGESFPGIYDALFDIESKVDPSKAWGEVKRQIYVA AFTVQAAAETLSEVA

“Recombinant” refers to polynucleotides, polypeptides, vectors, virusesand other macromolecules that are prepared, expressed, created orisolated by recombinant means.

“Regulatory element” refers to any cis- or trans acting genetic elementthat controls some aspect of the expression of nucleic acid sequences.

“Relapsed” refers to the return of a disease or the signs and symptomsof a disease after a period of improvement after prior treatment with atherapeutic.

“Refractory” refers to a disease that does not respond to a treatment. Arefractory disease can be resistant to a treatment before or at thebeginning of the treatment, or a refractory disease can become resistantduring a treatment.

“Single chain Fv” or “scFv” refers to a fusion protein comprising atleast one antibody fragment comprising a light chain variable region(VL) and at least one antibody fragment comprising a heavy chainvariable region (VH), wherein the VL and the VH are contiguously linkedvia a polypeptide linker, and capable of being expressed as a singlechain polypeptide. Unless specified, as used herein, a scFv may have theVL and VH variable regions in either order, e.g., with respect to theN-terminal and C-terminal ends of the polypeptide, the scFv may compriseVL-linker-VH or may comprise VH-linker-VL.

“(scFv)₂” or “tandem scFv” or “bis-scFv” fragments refers to a fusionprotein comprising two light chain variable region (VL) and two heavychain variable region (VH), wherein the two VL and the two VH arecontiguously linked via polypeptide linkers, and capable of beingexpressed as a single chain polypeptide. The two VL and two VH are fusedby peptide linkers to form a bivalent moleculeVL_(A)-linker-VH_(A)-linker-VL_(B)-linker-VH_(B) to form two bindingsites, capable of binding two different antigens or epitopesconcurrently.

“Specifically binds,” “specific binding,” “specifically binding” or“binds” refer to a proteinaceous molecule binding to an antigen or anepitope within the antigen with greater affinity than for otherantigens. Typically, the proteinaceous molecule binds to the antigen orthe epitope within the antigen with an equilibrium dissociation constant(K_(D)) of about 1×10⁻⁷ M or less, for example about 5×10⁻⁸ M or less,about 1×10⁻⁸ M or less, about 1×10⁻⁹ M or less, about 1×10⁻¹⁰ M or less,about 1×10⁻¹¹ M or less, or about 1×10⁻¹² M or less, typically with theK_(D) that is at least one hundred fold less than its K_(D) for bindingto a non-specific antigen (e.g., BSA, casein). In the context of theprostate neoantigens described here, “specific binding” refers tobinding of the proteinaceous molecule to the prostate neoantigen withoutdetectable binding to a wild-type protein the neoantigen is a variantof.

“Subject” includes any human or nonhuman animal. “Nonhuman animal”includes all vertebrates, e.g., mammals and non-mammals, such asnonhuman primates, sheep, dogs, cats, horses, cows, chickens,amphibians, reptiles, etc. The terms “subject” and “patient” can be usedinterchangeably herein.

“T cell” and “T lymphocyte” are interchangeable and used synonymouslyherein. T cell includes thymocytes, naïve T lymphocytes, memory T cells,immature T lymphocytes, mature T lymphocytes, resting T lymphocytes, oractivated T lymphocytes. A T cell can be a T helper (Th) cell, forexample a T helper 1 (Th1) or a T helper 2 (Th2) cell. The T cell can bea helper T cell (HTL; CD4⁺ T cell) CD4⁺ T cell, a cytotoxic T cell (CTL;CD8⁺ T cell), a tumor infiltrating cytotoxic T cell (TIL; CD8⁺ T cell),CD4⁺CD8⁺ T cell, or any other subset of T cells. Also included are “NKTcells”, which refer to a specialized population of T cells that expressa semi-invariant αβ T-cell receptor, but also express a variety ofmolecular markers that are typically associated with NK cells, such asNK1.1. NKT cells include NK1.1⁺ and NK1.1⁻, as well as CD4⁺, CD4⁻, CD8⁺and CD8⁻ cells. The TCR on NKT cells is unique in that it recognizesglycolipid antigens presented by the MHC I-like molecule CD Id. NKTcells can have either protective or deleterious effects due to theirabilities to produce cytokines that promote either inflammation orimmune tolerance. Also included are “gamma-delta T cells (γδ T cells),”which refer to a specialized population that to a small subset of Tcells possessing a distinct TCR on their surface, and unlike themajority of T cells in which the TCR is composed of two glycoproteinchains designated α- and β-TCR chains, the TCR in γδ T cells is made upof a γ-chain and a δ-chain. γδ T cells can play a role inimmunosurveillance and immunoregulation, and were found to be animportant source of IL-17 and to induce robust CD8⁺ cytotoxic T cellresponse. Also included are “regulatory T cells” or “Tregs” which referto T cells that suppress an abnormal or excessive immune response andplay a role in immune tolerance. Tregs are typically transcriptionfactor Foxp3-positive CD4⁺ T cells and can also include transcriptionfactor Foxp3-negative regulatory T cells that are IL-10-producing CD4⁺ Tcells.

“Therapeutically effective amount” or “effective amount” usedinterchangeably herein, refers to an amount effective, at dosages andfor periods of time necessary, to achieve a desired therapeutic result.A therapeutically effective amount may vary according to factors such asthe disease state, age, sex, and weight of the individual, and theability of a therapeutic or a combination of therapeutics to elicit adesired response in the individual. Example indicators of an effectivetherapeutic or combination of therapeutics that include, for example,improved wellbeing of the patient, reduction of a tumor burden, arrestedor slowed growth of a tumor, and/or absence of metastasis of cancercells to other locations in the body.

“Transduction” refers to the introduction of a foreign nucleic acid intoa cell using a viral vector.

“Treat,” “treating” or “treatment” of a disease or disorder such ascancer refers to accomplishing one or more of the following: reducingthe severity and/or duration of the disorder, inhibiting worsening ofsymptoms characteristic of the disorder being treated, limiting orpreventing recurrence of the disorder in subjects that have previouslyhad the disorder, or limiting or preventing recurrence of symptoms insubjects that were previously symptomatic for the disorder.

“Tumor cell” or a “cancer cell” refers to a cancerous, pre-cancerous ortransformed cell, either in vivo, ex vivo, or in tissue culture, thathas spontaneous or induced phenotypic changes. These changes do notnecessarily involve the uptake of new genetic material. Althoughtransformation may arise from infection with a transforming virus andincorporation of new genomic nucleic acid, uptake of exogenous nucleicacid or it can also arise spontaneously or following exposure to acarcinogen, thereby mutating an endogenous gene. Transformation/canceris exemplified by morphological changes, immortalization of cells,aberrant growth control, foci formation, proliferation, malignancy,modulation of tumor specific marker levels, invasiveness, tumor growthin suitable animal hosts such as nude mice, and the like, in vitro, invivo, and ex vivo.

“Variant,” “mutant” or “altered” refers to a polypeptide or apolynucleotide that differs from a reference polypeptide or a referencepolynucleotide by one or more modifications, for example one or moresubstitutions, insertions or deletions.

The numbering of amino acid residues in the antibody constant regionthroughout the specification is according to the EU index as describedin Kabat et al., Sequences of Proteins of Immunological Interest, 5thEd. Public Health Service, National Institutes of Health, Bethesda, Md.(1991), unless otherwise explicitly stated.

Mutations in the Ig constant regions are referred to as follows:L351Y_F405A_Y407V refers to L351Y, F405A and Y407V mutations in oneimmunoglobulin constant region. L351Y_F405A_Y407V/T394W refers to L351Y,F405A and Y407V mutations in the first Ig constant region and T394Wmutation in the second Ig constant region, which are present in onemultimeric protein.

“VHH” refers to a single-domain antibody or nanobody, exclusivelycomposed by heavy chain homodimers A VHH single domain antibody lack thelight chain and the CH1 domain of the heavy chain of conventional Fabregion.

Unless otherwise stated, any numerical values, such as a concentrationor a concentration range described herein, are to be understood as beingmodified in all instances by the term “about.” Thus, a numerical valuetypically includes ±10% of the recited value. For example, aconcentration of 1 mg/mL includes 0.9 mg/mL to 1.1 mg/mL. Likewise, aconcentration range of 1% to 10% (w/v) includes 0.9% (w/v) to 11% (w/v).As used herein, the use of a numerical range expressly includes allpossible subranges, all individual numerical values within that range,including integers within such ranges and fractions of the values unlessthe context clearly indicates otherwise.

The numbering of amino acid residues in the antibody constant regionthroughout the specification is according to the EU index as describedin Kabat et al., Sequences of Proteins of Immunological Interest, 5thEd. Public Health Service, National Institutes of Health, Bethesda, Md.(1991), unless otherwise explicitly stated.

TABLE 1 Conventional one- and three-letter amino acid codes used hereinAmino acid Three-letter code One-letter code Alanine Ala A Arginine ArgR Asparagine Asn N Aspartate Asp D Cysteine Cys C Glutamate Glu EGlutamine Gln Q Glycine Gly G Histidine His H Isoleucine Ile I LysineLys K Methionine Met M Phenylalanine Phe F Proline Pro P Serine Ser SThreonine Thr T Tryptophan Trp W Tyrosine Tyr Y Valine Val V

Antigen Binding Domains that Bind CD3ε.

The disclosure provides antigen binding domains that bind CD3ε,monospecific and multispecific proteins comprising the antigen bindingdomains that bind CD3ε, polynucleotides encoding the foregoing, vectors,host cells and methods of making and using the foregoing. The antigenbinding domains that bind CD3ε identified herein demonstrated severaladvantageous properties. First, the selection of IGHV1-69*02-IGHJ1-01and IGKV3-11*02-IGKJ4-01 germlines for CDR grafting ensured enhancedbinding as compared to the murine Cris-7 parent antibody. Second, uponintroducing human-to-mouse mutations, the selected clones demonstratedimproved thermostability by retaining binding after heat shock,including at 55° C., 60° C., and/or 65° C., a characteristic leading toimproved manufacturability and storage. This was not the case for themurine Cris-7 parent antibody, which demonstrated minimal binding at allto recombinant CD3 and T cells after heat shock when compared to antigenbinding domains that bind CD3ε of the present invention. Third, the PostTranslational Modification (PTM) risk was mitigated by substituting atposition N106 in SEQ ID Nos: 55, 54, and 48, and thus preventing Asndeamidation, which, if left unmodified, could lead to loss of activity.The engineered position at residue N106 was within HCDR3. Even withmutations at this position within HCR3, antibodies still retained theability to robustly bind antigen while also possessing added beneficialproperties (e.g., improved thermostability).

The disclosure also provides an isolated protein comprising an antigenbinding domain that binds CD3ε, wherein the antigen binding domain thatbinds CD3ε comprises the HCDR1, the HCDR2, the HCDR3, the LCDR1, theLCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 86, 79, 80, and 81,respectively. SEQ ID NO: 86 (PQVHYDYXGFPY, wherein X can be Q, A, G, orS) represents a genus HCDR3 amino acid sequence encompassing variantsdemonstrating improved properties, including improved thermostability,reduced deamidation risk and varied affinity to CD3, depending on theamino acid in place of “X”. For example, if X in SEQ ID NO: 86 issubstituted with either Q or A, the CD3 affinity is similar to theparental (having N in place of X); and if X is substituted with either Gor S, the CD3 affinity is lower compared to Q or A. This provided theadvantageous ability to tune the activity of T cell redirection abilityof the multi- or bi-specific proteins comprising the CD3ε biding domainsof the disclosure, in order to potentially mitigate cytokine response insubjects and potentially enhance tumor distribution of the multi- orbi-specific proteins.

The disclosure provides an isolated protein comprising an antigenbinding domain that binds CD3ε, wherein the antigen binding domain thatbinds CD3ε comprises:

-   -   a HCDR1, a HCDR2 and a HCDR3 of a heavy chain variable region        (VH) of SEQ ID NO: 55 and a light chain complementarity        determining region (LCDR) 1, a LCDR2 and a LCDR3 of a light        chain variable region (VL) of SEQ ID NO: 59;    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 55        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        58;    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 54        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        56; or    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 48        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        58;        -   wherein the amino acid in position N106 of SEQ ID NO: 55,            54, or 48 is optionally substituted with the amino acid            selected from the group consisting of A, G, S, F, E, T, R,            V, I, Y, L, P, Q, and K, wherein the residue numbering            starts from N-terminus of SEQ ID NO: 55, 54, or 48.

The disclosure provides an isolated protein comprising an antigenbinding domain that binds CD3ε, wherein the antigen binding domain thatbinds CD3ε comprises the HCDR1, the HCDR1, the HCDR3, the LCDR1, theLCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 86, 79, 80, and 81,respectively.

The disclosure provides an isolated protein comprising an antigenbinding domain that binds CD3ε, wherein the antigen binding domain thatbinds CD3ε comprises the HCDR1, the HCDR1, the HCDR3, the LCDR1, theLCDR2 and the LCDR3 of

SEQ ID NOs: 70, 71, 72, 79, 80, and 81, respectively;

SEQ ID NOs: 70, 71, 87, 79, 80, and 81, respectively; or

SEQ ID NOs: 70, 71, 90, 79, 80, and 81, respectively.

The disclosure provides an isolated protein comprising an antigenbinding domain that binds CD3ε, wherein the antigen binding domain thatbinds CD3ε comprises the VH of SEQ ID NOs: 55, 54, or 48 and the VL ofSEQ ID NOs: 59, 58, or 56.

The disclosure provides an isolated protein comprising an antigenbinding domain that binds CD3ε, wherein the antigen binding domain thatbinds CD3ε comprises

-   -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59;    -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 56; or    -   the VH of SEQ ID NO: 58 and the VL of SEQ ID NO: 58.

In other embodiments, the antigen binding domain that binds CD3ε is ascFv.

In other embodiments, the antigen binding domain that binds CD3ε is a(scFv)₂.

In other embodiments, the antigen binding domain that binds CD3ε is aFv.

In other embodiments, the antigen binding domain that binds CD3ε is aFab.

In other embodiments, the antigen binding domain that binds CD3ε is aF(ab′)2.

In other embodiments, the antigen binding domain that binds CD3ε is aFd.

In other embodiments, the CD3ε antigen binding domain is a dAb.

In other embodiments, the CD3ε antigen binding domain is a VHH.

CD3ε Binding scFvs

Any of the VH and the VL domains identified herein that bind CD3ε may beengineered into scFv format in either VH-linker-VL or VL-linker-VHorientation. Any of the VH and the VL domains identified herein may alsobe used to generate sc(Fv)₂ structures, such asVH-linker-VL-linker-VL-linker-VH, VH-linker-VL-linker-VH-linker-VL.VH-linker-VH-linker-VL-linker-VL. VL-linker-VH-linker-VH-linker-VL.VL-linker-VH-linker-VL-linker-VH or VL-linker-VL-linker-VH-linker-VH.

The VH and the VL domains identified herein may be incorporated into ascFv format and the binding and thermostability of the resulting scFv toCD3ε may be assessed using known methods. Binding may be assessed usingProteOn XPR36, Biacore 3000 or KinExA instrumentation, ELISA orcompetitive binding assays known to those skilled in the art. Bindingmay be evaluated using purified scFvs or E. coli supernatants or lysedcells containing the expressed scFv. The measured affinity of a testscFv to CD3ε may vary if measured under different conditions (e.g.,osmolarity, pH). Thus, measurements of affinity and other bindingparameters (e.g., KD, Kon, Koff) are typically made with standardizedconditions and standardized buffers. Thermostability may be evaluated byheating the test scFv at elevated temperatures, such as at 50° C., 55°C. or 60° C. for a period of time, such as 5 minutes (min), 10 min, 15min, 20 min, 25 min or 30 min and measuring binding of the test scFv toCD3ε. The scFvs retaining comparable binding to CD3ε when compared to anon-heated scFv sample are referred to as being thermostable.

In recombinant expression systems, the linker is a peptide linker andmay include any naturally occurring amino acid. Exemplary amino acidsthat may be included into the linker are Gly, Ser Pro, Thr, Glu, Lys,Arg, Ile, Leu, His and The. The linker should have a length that isadequate to link the VH and the VL in such a way that they form thecorrect conformation relative to one another so that they retain thedesired activity, such as binding to CD3ε.

The linker may be about 5-50 amino acids long. In other embodiments, thelinker is about 10-40 amino acids long. In other embodiments, the linkeris about 10-35 amino acids long. In other embodiments, the linker isabout 10-30 amino acids long. In other embodiments, the linker is about10-25 amino acids long. In other embodiments, the linker is about 10-20amino acids long. In other embodiments, the linker is about 15-20 aminoacids long. In other embodiments, the linker is about 16-19 amino acidslong. In other embodiments, the linker is 6 amino acids long. In otherembodiments, the linker is 7 amino acids long. In other embodiments, thelinker is 8 amino acids long. In other embodiments, the linker is 9amino acids long. In other embodiments, the linker is 10 amino acidslong. In other embodiments, the linker is 11 amino acids long. In otherembodiments, the linker is 12 amino acids long. In other embodiments,the linker is 13 amino acids long. In other embodiments, the linker is14 amino acids long. In other embodiments, the linker is 15 amino acidslong. In other embodiments, the linker is 16 amino acids long. In otherembodiments, the linker is 17 amino acids long. In other embodiments,the linker is 18 amino acids long. In other embodiments, the linker is19 amino acids long. In other embodiments, the linker is 20 amino acidslong. In other embodiments, the linker is 21 amino acids long. In otherembodiments, the linker is 22 amino acids long. In other embodiments,the linker is 23 amino acids long. In other embodiments, the linker is24 amino acids long. In other embodiments, the linker is 25 amino acidslong. In other embodiments, the linker is 26 amino acids long. In otherembodiments, the linker is 27 amino acids long. In other embodiments,the linker is 28 amino acids long. In other embodiments, the linker is29 amino acids long. In other embodiments, the linker is 30 amino acidslong. In other embodiments, the linker is 31 amino acids long. In otherembodiments, the linker is 32 amino acids long. In other embodiments,the linker is 33 amino acids long. In other embodiments, the linker is34 amino acids long. In other embodiments, the linker is 35 amino acidslong. In other embodiments, the linker is 36 amino acids long. In otherembodiments, the linker is 37 amino acids long. In other embodiments,the linker is 38 amino acids long. In other embodiments, the linker is39 amino acids long. In other embodiments, the linker is 40 amino acidslong. Exemplary linkers that may be used are Gly rich linkers, Gly andSer containing linkers, Gly and Ala containing linkers, Ala and Sercontaining linkers, and other flexible linkers.

Other linker sequences may include portions of immunoglobulin hingearea, CL or CH1 derived from any immunoglobulin heavy or light chainisotype. Alternatively, a variety of non-proteinaceous polymers,including polyethylene glycol (PEG), polypropylene glycol,polyoxyalkylenes, or copolymers of polyethylene glycol and polypropyleneglycol, may find use as linkers. Exemplary linkers that may be used areshown in Table 2. Additional linkers are described for example in Int.Pat. Publ. No. WO2019/060695.

TABLE 2 Linkers. Linker SEQ name Amino acid sequence ID NO: Linker 1GGSEGKSSGSGSESKSTGGS  3 Linker 2 GGGSGGGS  4 Linker 3 GGGSGGGSGGGS  5Linker 4 GGGSGGGSGGGSGGGS  6 Linker 5 GGGSGGGSGGGSGGGSGGGS  7 Linker 6GGGGSGGGGSGGGGS  8 Linker 7 GGGGSGGGGSGGGGSGGGGS  9 Linker 8GGGGSGGGGSGGGGSGGGGSGGGGS 10 Linker 9 GSTSGSGKPGSGEGSTKG 11 Linker 10IRPRAIGGSKPRVA 12 Linker 11 GKGGSGKGGSGKGGS 13 Linker 12 GGKGSGGKGSGGKGS14 Linker 13 GGGKSGGGKSGGGKS 15 Linker 14 GKGKSGKGKSGKGKS 16 Linker 15GGGKSGGKGSGKGGS 17 Linker 16 GKPGSGKPGSGKPGS 18 Linker 17GKPGSGKPGSGKPGSGKPGS 19 Linker 18 GKGKSGKGKSGKGKSGKGKS 20 Linker 19STAGDTHLGGEDFD 21 Linker 20 GEGGSGEGGSGEGGS 22 Linker 21 GGEGSGGEGSGGEGS23 Linker 22 GEGESGEGESGEGES 24 Linker 23 GGGESGGEGSGEGGS 25 Linker 24GEGESGEGESGEGESGEGES 26 Linker 25 GSTSGSGKPGSGEGSTKG 27 Linker 26PRGASKSGSASQTGSAPGS 28 Linker 27 GTAAAGAGAAGGAAAGAAG 29 Linker 28GTSGSSGSGSGGSGSGGGG 30 Linker 29 GKPGSGKPGSGKPGSGKPGS 31 Linker 30 GSGS32 Linker 31 APAPAPAPAP 33 Linker 32 APAPAPAPAPAPAPAPAPAP 34 Linker 33AEAAAKEAAAKEAAAAKEAAAAKEAAAA 35 KAAA Linker 34 GTEGKSSGSGSESKST 36

In other embodiments, the scFv comprises, from the N- to C-terminus, aVH, a first linker (L1) and a VL (VH-L1-VL).

In other embodiments, the scFv comprises, from the N-to C-terminus, theVL, the L1 and the VH (VL-L1-VH).

In other embodiments, the L1 comprises the amino acid sequence of 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or 36.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 3.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 4.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 5.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 6.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 7.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 8.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 9.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 10.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 11.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 12.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 13.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 14.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 15.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 16.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 17.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 17.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 19.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 20.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 21.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 22.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 23.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 24.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 25.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 26.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 27.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 28.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 29.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 30.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 31.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 32.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 33.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 34.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 35.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 36.

In other embodiments, the scFv comprises

-   -   a HCDR1, a HCDR2 and a HCDR3 of a heavy chain variable region        (VH) of SEQ ID NO: 55 and a light chain complementarity        determining region (LCDR) 1, a LCDR2 and a LCDR3 of a light        chain variable region (VL) of SEQ ID NO: 59;    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 55        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        58;    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 54        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        56; or    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 48        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        58;    -   wherein the amino acid in position N106 of SEQ ID NO: 55, 54, or        48 is optionally substituted with the amino acid selected from        the group consisting of A, G, S, F, E, T, R, V, I, Y, L, P, Q,        and K, wherein the residue numbering starts from N-terminus of        SEQ ID NO: 55, 54, or 48.

In other embodiments, the scFv comprises the HCDR1, the HCDR1, theHCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 86, 79,80, and 81, respectively.

In other embodiments, the scFv comprises the HCDR1, the HCDR1, theHCDR3, the LCDR1, the LCDR2 and the LCDR3 of

SEQ ID NOs: 70, 71, 72, 79, 80, and 81, respectively;

SEQ ID NOs: 70, 71, 87, 79, 80, and 81, respectively; or

SEQ ID NOs: 70, 71, 90, 79, 80, and 81, respectively.

In other embodiments, the scFv comprises the HCDR1, the HCDR1, theHCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 72, 79,80, and 81, respectively.

In other embodiments, the scFv comprises the HCDR1, the HCDR1, theHCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 87, 79,80, and 81, respectively.

In other embodiments, the scFv comprises the HCDR1, the HCDR1, theHCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 90, 79,80, and 81, respectively.

In other embodiments, the scFv comprises the VH of SEQ ID NOs: 55, 54,or 48 and the VL of SEQ ID NOs: 59, 58, or 56.

In other embodiments, the scFv comprises the VH of SEQ ID NO: 55 and theVL of SEQ ID NO: 59.

In other embodiments, the scFv comprises the VH of SEQ ID NO: 55 and theVL of SEQ ID NO: 58.

In other embodiments, the scFv comprises the VH of SEQ ID NO: 54 and theVL of SEQ ID NO: 56.

In other embodiments, the scFv comprises the VH of SEQ ID NO: 48 and theVL of SEQ ID NO: 58.

In other embodiments, the scFv comprises the VH of SEQ ID NO: 88 and theVL of SEQ ID NO: 58.

In other embodiments, the scFv comprises the VH of SEQ ID NO: 242 andthe VL of SEQ ID NO: 58.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NOs: 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108,109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122,123, 124, 125, or 126.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 96.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 97.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 98.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 99.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 100.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 101.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 102.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 103.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 104.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 105.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 106.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 107.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 108.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 109.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 110.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 111.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 112.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 113.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 114.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 115.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 116.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 117.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 118.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 119.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 120.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 121.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 122.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 123.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 124.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 125.

In other embodiments, the scFv comprises the amino acid sequence of SEQID NO: 126.

Other Antigen Binding Domains that Bind CD3ε

Any of the VH and the VL domains identified herein that bind CD3ε mayalso be engineered into Fab, F(ab′)₂, Fd or Fv format and their bindingto CD3ε and thermostability may be assessed using the assays describedherein. In certain embodiments thermostability is improved 2 fold, 3fold, 4 fold, 5 fold, upto 100 fold, with every integer in between (forexample, 6 fold, 7 fold, 8 fold, 9 fold, 10 fold, and so forth),compared to the murine Cris-7 parent antibody at 55° C., 60° C., and/or65° C. by the methods described herein.

In other embodiments, the Fab comprises

-   -   a HCDR1, a HCDR2 and a HCDR3 of a heavy chain variable region        (VH) of SEQ ID NO: 55 and a light chain complementarity        determining region (LCDR) 1, a LCDR2 and a LCDR3 of a light        chain variable region (VL) of SEQ ID NO: 59;    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 55        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        58;    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 54        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        56; or    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 48        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        58;

wherein the amino acid in position N106 of SEQ ID NO: 55, 54, or 48 isoptionally substituted with the amino acid selected from the groupconsisting of A, G, S, F, E, T, R, V, I, Y, L, P, Q, and K, wherein theresidue numbering starts from N-terminus of SEQ ID NO: 55, 54, or 48.

In other embodiments, the Fab comprises the HCDR1, the HCDR1, the HCDR3,the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 86, 79, 80,and 81, respectively.

In other embodiments, the Fab comprises the HCDR1, the HCDR1, the HCDR3,the LCDR1, the LCDR2 and the LCDR3 of

-   -   SEQ ID NOs: 70, 71, 72, 79, 80, and 81, respectively;    -   SEQ ID NOs: 70, 71, 87, 79, 80, and 81, respectively; or    -   SEQ ID NOs: 70, 71, 90, 79, 80, and 81, respectively.

In other embodiments, the Fab comprises the HCDR1, the HCDR2, the HCDR3,the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 72, 79, 80,and 81, respectively.

In other embodiments, the Fab comprises the HCDR1, the HCDR1, the HCDR3,the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 87, 79, 80,and 81, respectively.

In other embodiments, the Fab comprises the HCDR1, the HCDR1, the HCDR3,the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 90, 79, 80,and 81, respectively.

In other embodiments, the Fab comprises the VH of SEQ ID NO: 55 and theVL of SEQ ID NO: 59.

In other embodiments, the Fab comprises the VH of SEQ ID NO: 55 and theVL of SEQ ID NO: 58.

In other embodiments, the Fab comprises the VH of SEQ ID NO: 54 and theVL of SEQ ID NO: 56.

In other embodiments, the Fab comprises the VH of SEQ ID NO: 48 and theVL of SEQ ID NO: 58.

In other embodiments, the Fab comprises the VH of SEQ ID NO: 88 and theVL of SEQ ID NO: 58.

In other embodiments, the Fab comprises the VH of SEQ ID NO: 242 and theVL of SEQ ID NO: 58.

In other embodiments, the F(ab′)2 comprises

-   -   a HCDR1, a HCDR2 and a HCDR3 of a heavy chain variable region        (VH) of SEQ ID NO: 55 and a light chain complementarity        determining region (LCDR) 1, a LCDR2 and a LCDR3 of a light        chain variable region (VL) of SEQ ID NO: 59;

the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 55 and theLCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO: 58;

-   -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 54        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        56; or    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 48        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        58;

wherein the amino acid in position N106 of SEQ ID NO: 55, 54, or 48 isoptionally substituted with the amino acid selected from the groupconsisting of A, G, S, F, E, T, R, V, I, Y, L, P, Q, and K, wherein theresidue numbering starts from N-terminus of SEQ ID NO: 55, 54, or 48.

In other embodiments, the F(ab′)₂ comprises the HCDR1, the HCDR1, theHCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 86, 79,80, and 81, respectively.

In other embodiments, the F(ab′)2 comprises the HCDR1, the HCDR1, theHCDR3, the LCDR1, the LCDR2 and the LCDR3 of

-   -   SEQ ID NOs: 70, 71, 72, 79, 80, and 81, respectively;    -   SEQ ID NOs: 70, 71, 87, 79, 80, and 81, respectively; or    -   SEQ ID NOs: 70, 71, 90, 79, 80, and 81, respectively.

In other embodiments, the F(ab′)2 comprises the HCDR1, the HCDR2, theHCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 72, 79,80, and 81, respectively.

In other embodiments, the F(ab′)2 comprises the HCDR1, the HCDR1, theHCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 87, 79,80, and 81, respectively.

In other embodiments, the F(ab′)2 comprises the HCDR1, the HCDR1, theHCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 90, 79,80, and 81, respectively.

In other embodiments, the F(ab′)₂ comprises the VH of SEQ ID NO: 55 andthe VL of SEQ ID NO: 59.

In other embodiments, the F(ab′)₂ comprises the VH of SEQ ID NO: 55 andthe VL of SEQ ID NO: 58.

In other embodiments, the F(ab′)2 comprises the VH of SEQ ID NO: 54 andthe VL of SEQ ID NO: 56.

In other embodiments, the F(ab′)2 comprises the VH of SEQ ID NO: 48 andthe VL of SEQ ID NO: 58.

In other embodiments, the F(ab′)2 comprises the VH of SEQ ID NO: 88 andthe VL of SEQ ID NO: 58.

In other embodiments, the F(ab′)2 comprises the VH of SEQ ID NO: 242 andthe VL of SEQ ID NO: 58.

In other embodiments, the Fv comprises

-   -   a HCDR1, a HCDR2 and a HCDR3 of a heavy chain variable region        (VH) of SEQ ID NO: 55 and a light chain complementarity        determining region (LCDR) 1, a LCDR2 and a LCDR3 of a light        chain variable region (VL) of SEQ ID NO: 59;    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 55        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        58;    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 54        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        56; or    -   the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID NO: 48        and the LCDR1, the LCDR2 and the LCDR3 of the VL of SEQ ID NO:        58;

wherein the amino acid in position N106 of SEQ ID NO: 55, 54, or 48 isoptionally substituted with the amino acid selected from the groupconsisting of A, G, S, F, E, T, R, V, I, Y, L, P, Q, and K, wherein theresidue numbering starts from N-terminus of SEQ ID NO: 55, 54, or 48.

In other embodiments, the Fv comprises the HCDR1, the HCDR1, the HCDR3,the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 86, 79, 80,and 81, respectively.

In other embodiments, the Fv comprises the HCDR1, the HCDR1, the HCDR3,the LCDR1, the LCDR2 and the LCDR3 of

-   -   SEQ ID NOs: 70, 71, 72, 79, 80, and 81, respectively;    -   SEQ ID NOs: 70, 71, 87, 79, 80, and 81, respectively; or    -   SEQ ID NOs: 70, 71, 90, 79, 80, and 81, respectively.

In other embodiments, the Fv comprises the HCDR1, the HCDR2, the HCDR3,the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 72, 79, 80,and 81, respectively.

In other embodiments, the Fv comprises the HCDR1, the HCDR1, the HCDR3,the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 87, 79, 80,and 81, respectively.

In other embodiments, the Fv comprises the HCDR1, the HCDR1, the HCDR3,the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71, 90, 79, 80,and 81, respectively.

In other embodiments, the Fv comprises the VH of SEQ ID NO: 55 and theVL of SEQ ID NO: 59.

In other embodiments, the Fv comprises the VH of SEQ ID NO: 55 and theVL of SEQ ID NO: 58.

In other embodiments, the Fv comprises the VH of SEQ ID NO: 54 and theVL of SEQ ID NO: 56.

In other embodiments, the Fv comprises the VH of SEQ ID NO: 48 and theVL of SEQ ID NO: 58.

In other embodiments, the Fv comprises the VH of SEQ ID NO: 88 and theVL of SEQ ID NO: 58.

In other embodiments, the Fv comprises the VH of SEQ ID NO: 242 and theVL of SEQ ID NO: 58.

In other embodiments, the Fd comprises a heavy chain complementaritydetermining region (HCDR) 1, a HCDR2 and a HCDR3 of a heavy chainvariable region (VH) of SEQ ID NOs: 55, 54, or 48.

In other embodiments, the Fd comprises the HCDR1, the HCDR2, and theHCDR3 of SEQ ID NOs: 70, 71, and 86, respectively.

In other embodiments, the Fd comprises the HCDR1, the HCDR1, and theHCDR3 of SEQ ID NOs: 70, 71, and 72, respectively.

In other embodiments, the Fd comprises the HCDR1, the HCDR1, and theHCDR3 of SEQ ID NOs: 70, 71, and 87, respectively.

In other embodiments, the Fd comprises the HCDR1, the HCDR1, and theHCDR3 of SEQ ID NOs: 70, 71, and 90, respectively.

In other embodiments, the Fd comprises the VH of SEQ ID NO: 55.

In other embodiments, the Fd comprises the VH of SEQ ID NO: 54.

In other embodiments, the Fd comprises the VH of SEQ ID NO: 48.

In other embodiments, the Fd comprises the VH of SEQ ID NO: 88.

In other embodiments, the Fd comprises the VH of SEQ ID NO: 242.

Homologous Antigen Binding Domains and Antigen Binding Domains withConservative Substitutions

Variants of the antigen binding domains that bind CD3ε are within thescope of the disclosure. For example, variants may comprise 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28 or 29 amino acid substitutions in the antigen bindingdomain that bind CD3ε as long as they retain or have improved functionalproperties when compared to the parent antigen binding domains. In otherembodiments, the sequence identity may be about 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or99% to the antigen binding domains that bind CD3ε of the disclosure. Inother embodiments, the variation is in the framework regions. In otherembodiments, variants are generated by conservative substitutions.

For example, the antigen binding domains that bind CD3ε may comprisesubstitutions at residue position N106 (residue numbering fromN-terminus of SEQ ID NO: 55, 54, or 48). Conservative substitutions maybe made at any indicated positions and the resulting variant antigenbinding domains that bind CD3ε are tested for their desiredcharacteristics in the assays described herein.

Also provided are antigen binding domains that bind CD3ε comprising theVH and the VL which are at least 80% identical to

-   -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59;    -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 56;    -   the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 88 and the VL of SEQ ID NO: 58; or    -   the VH of SEQ ID NO: 242 and the VL of SEQ ID NO: 58.

In other embodiments, the identity is 85%. In other embodiments, theidentity is 90%. In other embodiments, the identity is 91%. In otherembodiments, the identity is 91%. In other embodiments, the identity is92%. In other embodiments, the identity is 93%. In other embodiments,the identity is 94%. In other embodiments, the identity is 94%. In otherembodiments, the identity is 95%. In other embodiments, the identity is96%. In other embodiments, the identity is 97%. In other embodiments,the identity is 98%. In other embodiments, the identity is 99%.

The percent identity between the two sequences is a function of thenumber of identical positions shared by the sequences (i.e., %identity=number of identical positions/total number of positions ×100),taking into account the number of gaps, and the length of each gap,which need to be introduced for optimal alignment of the two sequences.

The percent identity between two amino acid sequences may be determinedusing the algorithm of E. Meyers and W. Miller (Comput Appl Biosci4:11-17 (1988)) which has been incorporated into the ALIGN program(version 2.0), using a PAM120 weight residue table, a gap length penaltyof 12 and a gap penalty of 4. In addition, the percent identity betweentwo amino acid sequences may be determined using the Needleman andWunsch (J Mol Biol 48:444-453 (1970)) algorithm which has beenincorporated into the GAP program in the GCG software package (can beretrieved from the Internet <URL: http://www.gcg.com>), using either aBlossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12,10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.

In other embodiments, variant antigen binding domains that bind CD3εcomprise one or two conservative substitutions in any of the CDRregions, while retaining desired functional properties of the parentantigen binding fragments that bind CD3ε.

“Conservative modifications” refer to amino acid modifications that donot significantly affect or alter the binding characteristics of theantibody containing the amino acid modifications. Conservativemodifications include amino acid substitutions, additions and deletions.Conservative amino acid substitutions are those in which the amino acidis replaced with an amino acid residue having a similar side chain. Thefamilies of amino acid residues having similar side chains are welldefined and include amino acids with acidic side chains (e.g., asparticacid, glutamic acid), basic side chains (e.g., lysine, arginine,histidine), nonpolar side chains (e.g., alanine, valine, leucine,isoleucine, proline, phenylalanine, methionine), uncharged polar sidechains (e.g., glycine, asparagine, glutamine, cysteine, serine,threonine, tyrosine, tryptophan), aromatic side chains (e.g.,phenylalanine, tryptophan, histidine, tyrosine), aliphatic side chains(e.g., glycine, alanine, valine, leucine, isoleucine, serine,threonine), amide (e.g., asparagine, glutamine), beta-branched sidechains (e.g., threonine, valine, isoleucine) and sulfur-containing sidechains (cysteine, methionine). Furthermore, any native residue in thepolypeptide may also be substituted with alanine, as has been previouslydescribed for alanine scanning mutagenesis (MacLennan et al., (1988)Acta Physiol Scand Suppl 643:55-67; Sasaki et al., (1988) Adv Biophys35:1-24). Amino acid substitutions to the antibodies of the inventionmay be made by known methods for example by PCR mutagenesis (U.S. Pat.No. 4,683,195). Alternatively, libraries of variants may be generatedfor example using random (NNK) or non-random codons, for example DVKcodons, which encode 11 amino acids (Ala, Cys, Asp, Glu, Gly, Lys, Asn,Arg, Ser, Tyr, Trp). The resulting variants may be tested for theircharacteristics using assays described herein.

Methods of Generating Antigen Binding Fragment that Bind CD3ε

Antigen binding domains that bind CD3ε provided in the disclosure may begenerated using various technologies. For example, the hybridoma methodof Kohler and Milstein may be used to identify VH/VL pairs that bindCD3ε. In the hybridoma method, a mouse or other host animal, such as ahamster, rat or chicken is immunized with human and/or cyno CD3ε,followed by fusion of spleen cells from immunized animals with myelomacells using standard methods to form hybridoma cells. Colonies arisingfrom single immortalized hybridoma cells may be screened for productionof the antibodies containing the antigen binding domains that bind CD3εwith desired properties, such as specificity of binding,cross-reactivity or lack thereof, affinity for the antigen, and anydesired functionality.

Antigen binding domains that bind CD3ε generated by immunizing non-humananimals may be humanized. Exemplary humanization techniques includingselection of human acceptor frameworks include CDR grafting (U.S. Pat.No. 5,225,539), SDR grafting (U.S. Pat. No. 6,818,749), Resurfacing(Padlan, (1991) Mol Immunol 28:489-499), Specificity DeterminingResidues Resurfacing (U.S. Patent Publ. No. 2010/0261620), humanframework adaptation (U.S. Pat. No. 8,748,356) or superhumanization(U.S. Pat. No. 7,709,226). In these methods, CDRs or a subset of CDRresidues of parental antibodies are transferred onto human frameworksthat may be selected based on their overall homology to the parentalframeworks, based on similarity in CDR length, or canonical structureidentity, or a combination thereof.

Humanized antigen biding domains may be further optimized to improvetheir selectivity or affinity to a desired antigen by incorporatingaltered framework support residues to preserve binding affinity(backmutations) by techniques such as those described in Int. PatentPubl. Nos. WO1090/007861 and WO1992/22653, or by introducing variationat any of the CDRs for example to improve affinity of the antigenbinding domain.

Transgenic animals, such as mice, rat or chicken carrying humanimmunoglobulin (Ig) loci in their genome may be used to generate antigenbinding fragments that bind CD3ε, and are described in for example U.S.Pat. No. 6,150,584, Int. Patent Publ. No. WO1999/45962, Int. PatentPubl. Nos. WO2002/066630, WO2002/43478, WO2002/043478 and WO1990/04036.The endogenous immunoglobulin loci in such animal may be disrupted ordeleted, and at least one complete or partial human immunoglobulin locusmay be inserted into the genome of the animal using homologous ornon-homologous recombination, using transchromosomes, or usingminigenes. Companies such as Regeneron (<URL:http://www.regeneron.com>), Harbour Antibodies(http://www.harbourantibodies.com), Open Monoclonal Technology, Inc.(OMT) (<URL: http://www.omtinc.net>), KyMab (<URL:http://www.kymab.com>), Trianni (<URL: http://www.trianni.com>) andAblexis (<URL: http://www.ablexis.com>) may be engaged to provide humanantibodies directed against a selected antigen using technologies asdescribed above.

Antigen binding domains that bind CD3ε may be selected from a phagedisplay library, where the phage is engineered to express humanimmunoglobulins or portions thereof such as Fabs, single chainantibodies (scFv), or unpaired or paired antibody variable regions. Theantigen binding domains that bind CD3ε may be isolated for example fromphage display library expressing antibody heavy and light chain variableregions as fusion proteins with bacteriophage pIX coat protein asdescribed in Shi et al., (2010) J Mol Biol 397:385-96, and Int. PatentPubl. No. WO09/085462). The libraries may be screened for phage bindingto human and/or cyno CD3ε and the obtained positive clones may befurther characterized, the Fabs isolated from the clone lysates, andconverted to scFvs or other configurations of antigen binding fragments.

Preparation of immunogenic antigens and expression and production ofantigen binding domains of the disclosure may be performed using anysuitable technique, such as recombinant protein production. Theimmunogenic antigens may be administered to an animal in the form ofpurified protein, or protein mixtures including whole cells or cell ortissue extracts, or the antigen may be formed de novo in the animal'sbody from nucleic acids encoding said antigen or a portion thereof.

Conjugation to Half-Life Extending Moieties

The antigen binding domains that bind CD3ε of the disclosure may beconjugated to a half-life extending moiety. Exemplary half-lifeextending moieties are albumin, albumin variants, albumin-bindingproteins and/or domains, transferrin and fragments and analoguesthereof, immunoglobulins (Ig) or fragments thereof, such as Fc regions.Amino acid sequences of the aforementioned half-life extending moietiesare known. Ig or fragments thereof include all isotypes (i.e., IgG1,IgG2, IgG3, IgG4, IgM, IgA and IgE).

Additional half-life extending moieties that may be conjugated to theantigen binding domains that bind CD3ε of the disclosure includepolyethylene glycol (PEG) molecules, such as PEG5000 or PEG20,000, fattyacids and fatty acid esters of different chain lengths, for examplelaurate, myristate, stearate, arachidate, behenate, oleate,arachidonate, octanedioic acid, tetradecanedioic acid, octadecanedioicacid, docosanedioic acid, and the like, polylysine, octane,carbohydrates (dextran, cellulose, oligo- or polysaccharides) fordesired properties. These moieties may be direct fusions with theantigen binding domains that bind CD3ε of the disclosure and may begenerated by standard cloning and expression techniques. Alternatively,well known chemical coupling methods may be used to attach the moietiesto recombinantly produced antigen binding domains that bind CD3ε of thedisclosure.

A pegyl moiety may for example be conjugated to the antigen bindingdomain that bind CD3ε of the disclosure by incorporating a cysteineresidue to the C-terminus of the antigen binding domain that bind CD3εof the disclosure, or engineering cysteines into residue positions thatface away from the CD3ε binding site and attaching a pegyl group to thecysteine using well known methods.

In other embodiments, the antigen binding fragment that binds CD3ε isconjugated to a half-life extending moiety.

In other embodiments, the half-life extending moiety is animmunoglobulin (Ig), a fragment of the Ig, an Ig constant region, afragment of the Ig constant region, a Fc region, transferrin, albumin,an albumin binding domain or polyethylene glycol. In other embodiments,the half-life extending moiety is an Ig constant region.

In other embodiments, the half-life extending moiety is the Ig.

In other embodiments, the half-life extending moiety is the fragment ofthe Ig.

In other embodiments, the half-life extending moiety is the Ig constantregion.

In other embodiments, the half-life extending moiety is the fragment ofthe Ig constant region.

In other embodiments, the half-life extending moiety is the Fc region.

In other embodiments, the half-life extending moiety is albumin.

In other embodiments, the half-life extending moiety is the albuminbinding domain.

In other embodiments, the half-life extending moiety is transferrin.

In other embodiments, the half-life extending moiety is polyethyleneglycol.

The antigen binding domains that bind CD3ε conjugated to a half-lifeextending moiety may be evaluated for their pharmacokinetic propertiesutilizing known in vivo models.

Conjugation to Immunoglobulin (Ig) Constant Regions or Fragments of theIg Constant Regions

The antigen binding domains that bind CD3ε of the disclosure may beconjugated to an Ig constant region or a fragment of the Ig constantregion to impart antibody-like properties, including Fc effectorfunctions C1q binding, complement dependent cytotoxicity (CDC), Fcreceptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC),phagocytosis or down regulation of cell surface receptors (e.g., B cellreceptor; BCR). The Ig constant region or the fragment of the Igconstant region functions also as a half-life extending moiety asdiscussed herein. The antigen binding domains that bind CD3ε of thedisclosure may be engineered into conventional full-length antibodiesusing standard methods. The full-length antibodies comprising theantigen binding domain that binds CD3ε may further be engineered asdescribed herein.

Immunoglobulin heavy chain constant region comprised of subdomains CH1,hinge, CH2 and CH3. The CH1 domain spans residues A118-V215, the CH2domain residues A231-K340 and the CH3 domain residues G341-K447 on theheavy chain, residue numbering according to the EU Index. In someinstances, G341 is referred as a CH2 domain residue. Hinge is generallydefined as including E216 and terminating at P230 of human IgG1. Ig Fcregion comprises at least the CH2 and the CH3 domains of the Ig constantregion, and therefore comprises at least a region from about A231 toK447 of Ig heavy chain constant region.

The invention also provides an antigen binding domain that binds CD3εconjugated to an immunoglobulin (Ig) constant region or a fragment ofthe Ig constant region.

In other embodiments, the Ig constant region is a heavy chain constantregion

In other embodiments, the Ig constant region is a light chain constantregion.

In other embodiments, the fragment of the Ig constant region comprises aFc region.

In other embodiments, the fragment of the Ig constant region comprises aCH2 domain.

In other embodiments, the fragment of the Ig constant region comprises aCH3 domain.

In other embodiments, the fragment of the Ig constant region comprisesthe CH2 domain and the CH3 domain.

In other embodiments, the fragment of the Ig constant region comprisesat least portion of a hinge, the CH2 domain and the CH3 domain. Portionof the hinge refers to one or more amino acid residues of the Ig hinge.

In other embodiments, the fragment of the Ig constant region comprisesthe hinge, the CH2 domain and the CH3 domain.

In other embodiments, the antigen binding domain that binds CD3ε isconjugated to the N-terminus of the Ig constant region or the fragmentof the Ig constant region.

In other embodiments, the antigen binding domain that binds CD3ε isconjugated to the C-terminus of the Ig constant region or the fragmentof the Ig constant region.

In other embodiments, the antigen binding domain that binds CD3ε isconjugated to the Ig constant region or the fragment of the Ig constantregion via a second linker (L2).

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNOs: 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or 36.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 3.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 4.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 5.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 6.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 7.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 8.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 9.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 10.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 11.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 12.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 13.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 14.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 15.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 16.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 17.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 17.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 19.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 20.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 21.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 22.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 23.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 24.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 25.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 26.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 27.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 28.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 29.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 30.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 31.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 32.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 33.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 34.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 35.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNO: 36.

The antigen binding domains that bind CD3ε of the disclosure conjugatedto Ig constant region or the fragment of the Ig constant region may beassessed for their functionality using several known assays. Binding toCD3ε may be assessed using methods described herein. Altered propertiesimparted by the Ig constant domain or the fragment of the Ig constantregion such as Fc region may be assayed in Fc receptor binding assaysusing soluble forms of the receptors, such as the FcγRI, FcγRII, FcγRIIIor FcRn receptors, or using cell-based assays measuring for exampleADCC, CDC or ADCP.

ADCC may be assessed using an in vitro assay using CD3ε expressing cellsas target cells and NK cells as effector cells. Cytolysis may bedetected by the release of label (e.g. radioactive substrates,fluorescent dyes or natural intracellular proteins) from the lysedcells. In an exemplary assay, target cells are used with a ratio of 1target cell to 4 effector cells. Target cells are pre-labeled with BATDAand combined with effector cells and the test antibody. The samples areincubated for 2 hours and cell lysis measured by measuring releasedBATDA into the supernatant. Data is normalized to maximal cytotoxicitywith 0.67% Triton X-100 (Sigma Aldrich) and minimal control determinedby spontaneous release of BATDA from target cells in the absence of anyantibody.

ADCP may be evaluated by using monocyte-derived macrophages as effectorcells and any CD3ε expressing cells as target cells which are engineeredto express GFP or other labeled molecule. In an exemplary assay,effector:target cell ratio may be for example 4:1. Effector cells may beincubated with target cells for 4 hours with or without the antibody ofthe invention. After incubation, cells may be detached using accutase.Macrophages may be identified with anti-CD11b and anti-CD14 antibodiescoupled to a fluorescent label, and percent phagocytosis may bedetermined based on % GFP fluorescence in the CD11⁺CD14⁺ macrophagesusing standard methods.

CDC of cells may be measured for example by plating Daudi cells at 1×10⁵cells/well (50 μL/well) in RPMI-B (RPMI supplemented with 1% BSA),adding 50 μL of test protein to the wells at final concentration between0-100 μg/mL, incubating the reaction for 15 min at room temperature,adding 11 μL of pooled human serum to the wells, and incubation thereaction for 45 min at 37° C. Percentage (%) lysed cells may be detectedas % propidium iodide stained cells in FACS assay using standardmethods.

Proteins Comprising the Antigen Binding Domains that Bind CD3ε of theDisclosure

The antigen binding domains that bind CD3ε of the disclosure may beengineered into monospecific or multispecific proteins of variousdesigns using standard methods.

The disclosure also provides a monospecific protein comprising theantigen binding domain that binds CD3ε of the disclosure.

In other embodiments, the monospecific protein is an antibody.

The disclosure also provides a multispecific protein comprising theantigen binding domain that binds CD3ε of the disclosure.

In other embodiments, the multispecific protein is bispecific.

In other embodiments, the multispecific protein is trispecific.

In other embodiments, the multispecific protein is tetraspecific.

In other embodiments, the multispecific protein is monovalent forbinding to CD3ε.

In other embodiments, the multispecific protein is bivalent for bindingto CD3ε.

The disclosure also provides an isolated multispecific proteincomprising a first antigen binding domain that binds CD3ε and a secondantigen binding domain that binds a tumor antigen. In other embodiments,the tumor antigen is a protein or a fragment thereof that is present ona cancer cell or specific to a cancer cell.

In other embodiments, the tumor antigen is a BCMA antigen. In otherembodiments, the tumor antigen is a PSMA antigen. In other embodiments,the tumor antigen is a CD79b antigen. In other embodiments, the tumorantigen is a CD20 antigen. In other embodiments, the tumor antigen is aCD20 antigen and a CD79b antigen.

In other embodiments, the first antigen binding domain that binds CD3εand/or the second antigen binding domain that binds the tumor antigencomprise a scFv, a (scFv)₂, a Fv, a Fab, a F(ab′)₂, a Fd, a dAb or a WM.

In other embodiments, the first antigen binding domain that binds CD3εand/or the second antigen binding domain that binds the tumor antigencomprise the Fab.

In other embodiments, the first antigen binding domain that binds CD3εand/or the second antigen binding domain that binds the tumor antigencomprise the F(ab′)2.

In other embodiments, the first antigen binding domain that binds CD3εand/or the second antigen binding domain that binds the tumor antigencomprise the VHH.

In other embodiments, the first antigen binding domain that binds CD3εand/or the second antigen binding domain that binds the tumor antigencomprise the Fv.

In other embodiments, the first antigen binding domain that binds CD3εand/or the second antigen binding domain that binds the tumor antigencomprise the Fd.

In other embodiments, the first antigen binding domain that binds CD3εand/or the second antigen binding domain that binds the tumor antigencomprise the scFv.

In other embodiments, the scFv comprises, from the N- to C-terminus, aVH, a first linker (L1) and a VL (VH-L1-VL) or the VL, the L1 and the VH(VL-L1-VH).

In other embodiments, the L1 comprises about 5-50 amino acids.

In other embodiments, the L1 comprises about 5-40 amino acids.

In other embodiments, the L1 comprises about 10-30 amino acids.

In other embodiments, the L1 comprises about 10-20 amino acids.

In other embodiments, the L1 comprises the amino acid sequence of 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or 36.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 3.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 4.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 5.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 6.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 7.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 8.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 9.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 10.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 11.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 12.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 13.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 14.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 15.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 16.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 17.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 17.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 19.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 20.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 21.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 22.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 23.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 24.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 25.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 26.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 27.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 28.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 29.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 30.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 31.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 32.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 33.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 34.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 35.

In other embodiments, the L1 comprises the amino acid sequence of SEQ IDNO: 36.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the HCDR1 of SEQ ID NO: 70, the HCDR2 of SEQ ID NO: 71, theHCDR3 of SEQ ID NOs: 72, 87, 90, or 86, the LCDR1 of SEQ ID NO: 79, theLCDR2 of SEQ ID NO: 80, and the LCDR3 of SEQ ID NOs: 81.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and theLCDR3 of SEQ ID NOs: 70, 71, 86, 79, 80, and 81, respectively.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and theLCDR3 of

-   -   SEQ ID NOs: 70, 71, 72, 79, 80, and 81, respectively;    -   SEQ ID NOs: 70, 71, 87, 79, 80, and 81, respectively; or    -   SEQ ID NOs: 70, 71, 90, 79, 80, and 81, respectively.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 56.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 58.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the VH of SEQ ID NO: 88 and the VL of SEQ ID NO: 58.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the VH of SEQ ID NO: 242 and the VL of SEQ ID NO: 58.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the VH of SEQ ID NOs: 55, 54, or 48 and the VL of SEQ ID NOs:59, 58, or 56.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID Nos: 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114,115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, or 126.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 96.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 97.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 98.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 99.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 100.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 101.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 102.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 103.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 104.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 105.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 106.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 107.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 108.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 109.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 110.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 112.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 113.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 114.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 115.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 116.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 117.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 118.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 119.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 120.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 121.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 122.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 123.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 124.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 125.

In other embodiments, the first antigen binding domain that binds CD3εcomprises the amino acid sequence of SEQ ID NO: 126.

In other embodiments, the second antigen binding domain that binds atumor antigen is specific to PSMA.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1242 and the LC of SEQ IDNO: 1243.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1244 and the LC of SEQ IDNO: 1245.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1246 and the LC of SEQ IDNO: 1247.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1248.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1250.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1252.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1254.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1256.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1258.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1260.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1262.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1264.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1266.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1268.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1270.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1272.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1274.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1276.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1278.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1280.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1282.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1284.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1286.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1288.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1290.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1292.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1294.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1459 and the LC of SEQ IDNO: 1460.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1461 and the LC of SEQ IDNO: 1462.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1356 and the LC of SEQ IDNO: 1357.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1358 and the LC of SEQ IDNO: 1359.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1360 and the LC of SEQ IDNO: 1361.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1362 and the LC of SEQ IDNO: 1363.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1364 and the LC of SEQ IDNO: 1365.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1366 and the LC of SEQ IDNO: 1367.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1368 and the LC of SEQ IDNO: 1369.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1370 and the LC of SEQ IDNO: 1371.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1372 and the LC of SEQ IDNO: 1373.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1374 and the LC of SEQ IDNO: 1375.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1376 and the LC of SEQ IDNO: 1377.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1378 and the LC of SEQ IDNO: 1379.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1378 and the LC of SEQ IDNO: 1379.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1380 and the LC of SEQ IDNO: 1381.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1382 and the LC of SEQ IDNO: 1383.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1384 and the LC of SEQ IDNO: 1385.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1386 and the LC of SEQ IDNO: 1387.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1388 and the LC of SEQ IDNO: 1389.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1390 and the LC of SEQ IDNO: 1391.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1392 and the LC of SEQ IDNO: 1393.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1394 and the LC of SEQ IDNO: 1395.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1396 and the LC of SEQ IDNO: 1397.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1396 and the LC of SEQ IDNO: 1397.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1396 and the LC of SEQ IDNO: 1397.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1396 and the LC of SEQ IDNO: 1397.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1398 and the LC of SEQ IDNO: 1399.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1400 and the LC of SEQ IDNO: 1401.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1402 and the LC of SEQ IDNO: 1403.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1404 and the LC of SEQ IDNO: 1405.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1406 and the LC of SEQ IDNO: 1407.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1408 and the LC of SEQ IDNO: 1409.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1410 and the LC of SEQ IDNO: 1411.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1412 and the LC of SEQ IDNO: 1413.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1414 and the LC of SEQ IDNO: 1415.

In other embodiments, the second antigen binding domain that binds atumor antigen is specific to CD79b.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1489 and the LC of SEQ IDNO: 1491.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1493 and the LC of SEQ IDNO: 1495.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1497 and the LC of SEQ IDNO: 1499.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1502 and the LC of SEQ IDNO: 1499.

In other embodiments, the second antigen binding domain that binds atumor antigen comprises the HC of SEQ ID NO: 1489 and the LC of SEQ IDNO: 1491.

In other embodiments, the first antigen binding domain that binds CD3εis conjugated to a first immunoglobulin (Ig) constant region or afragment of the first Ig constant region and/or the second antigenbinding domain that binds the tumor antigen is conjugated to a secondimmunoglobulin (Ig) constant region or a fragment of the second Igconstant region.

In other embodiments, the fragment of the first Ig constant regionand/or the fragment of the second Ig constant region comprises a Fcregion.

In other embodiments, the fragment of the first Ig constant regionand/or the fragment of the second Ig constant region comprises a CH2domain.

In other embodiments, the fragment of the first Ig constant regionand/or the fragment of the second Ig constant region comprises a CH3domain.

In other embodiments, the fragment of the first Ig constant regionand/or the fragment of the second Ig constant region comprises the CH2domain and the CH3 domain.

In other embodiments, the fragment of the first Ig constant regionand/or the fragment of the second Ig constant region comprises at leastportion of a hinge, the CH2 domain and the CH3 domain.

In other embodiments, the fragment of the Ig constant region comprisesthe hinge, the CH2 domain and the CH3 domain.

In other embodiments, the multispecific protein further comprises asecond linker (L2) between the first antigen binding domain that bindsCD3ε and the first Ig constant region or the fragment of the first Igconstant region and the second antigen binding domain that binds thetumor antigen and the second Ig constant region or the fragment of thesecond Ig constant region.

In other embodiments, the L2 comprises the amino acid sequence of SEQ IDNOs: 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or 36.

In other embodiments, the first Ig constant region or the fragment ofthe first Ig constant region and the second Ig constant region or thefragment of the second Ig constant region is an IgG1, an IgG2, and IgG3or an IgG4 isotype.

In other embodiments, the first Ig constant region or the fragment ofthe first Ig constant region and the second Ig constant region or thefragment of the second Ig constant region is an IgG1 isotype.

In other embodiments, the first Ig constant region or the fragment ofthe first Ig constant region and the second Ig constant region or thefragment of the second Ig constant region is an IgG2 isotype.

In other embodiments, the first Ig constant region or the fragment ofthe first Ig constant region and the second Ig constant region or thefragment of the second Ig constant region is an IgG3 isotype.

In other embodiments, the first Ig constant region or the fragment ofthe first Ig constant region and the second Ig constant region or thefragment of the second Ig constant region is an IgG4 isotype.

The first Ig constant region or the fragment of the first Ig constantregion and the second Ig constant region or the fragment of the secondIg constant region can further be engineered as described herein.

In other embodiments, the first Ig constant region or the fragment ofthe first Ig constant region and the second Ig constant region or thefragment of the second Ig constant region comprises at least onemutation that results in reduced binding of the multispecific protein toa FcγR.

In other embodiments, the at least one mutation that results in reducedbinding of the multispecific protein to the FcγR is selected from thegroup consisting of F234A/L235A, L234A/L235A, L234A/L235A/D265S,V234A/G237A/P238S/H268A/V309L/A330S/P331S, F234A/L235A,S228P/F234A/L235A, N297A, V234A/G237A,K214T/E233P/L234V/L235A/G236-deleted/A327G/P331A/D365E/L358M,H268Q/V309L/A330S/P331S, S267E/L328F, L234F/L235E/D265A,L234A/L235A/G237A/P238S/H268A/A330S/P331S, S228P/F234A/L235A/G237A/P238Sand S228P/F234A/L235A/G236-deleted/G237A/P238S, wherein residuenumbering is according to the EU index.

In other embodiments, the first Ig constant region or the fragment ofthe first Ig constant region and the second Ig constant region or thefragment of the second Ig constant region comprises at least onemutation that results in enhanced binding of the multispecific proteinto a Fcγ receptor (FcγR).

In other embodiments, the at least one mutation that results in enhancedbinding of the multispecific protein to the FcγR is selected from thegroup consisting of S239D/I332E, S298A/E333A/K334A, F243L/R292P/Y300L,F243L/R292P/Y300L/P396L, F243L/R292P/Y300L/V305I/P396L andG236A/S239D/I332E, wherein residue numbering is according to the EUindex.

In other embodiments, the FcγR is FcγRI, FcγRIIA, FcγRIIB or FcγRIII, orany combination thereof.

In other embodiments, the first Ig constant region or the fragment ofthe first Ig constant region and the second Ig constant region or thefragment of the second Ig constant region comprises at least onemutation that modulates a half-life of the multispecific protein.

In other embodiments, the at least one mutation that modulates thehalf-life of the multispecific protein is selected from the groupconsisting of H435A, P257I/N434H, D376V/N434H,M252Y/S254T/T256E/H433K/N434F, T308P/N434A and H435R, wherein residuenumbering is according to the EU index.

In other embodiments, the multispecific protein comprises at least onemutation in a CH3 domain of the first Ig constant region or in a CH3domain of the fragment of the first Ig constant region and/or at leastone mutation in a CH3 domain of the second Ig constant region or in aCH3 domain of the fragment of the second Ig constant region.

In other embodiments, the at least one mutation in a CH3 domain of thefirst Ig constant region or in a CH3 domain of the fragment of the firstIg constant region and/or at least one mutation in a CH3 domain of thesecond Ig constant region or in a CH3 domain of the fragment of thesecond Ig constant region is selected from the group consisting ofT350V, L351Y, F405A, Y407V, T366Y, T366W, T366L, T366L, F405W, T394W,K392L, T394S, T394W, Y407T, Y407A, T366S/L368A/Y407V, L351Y/F405A/Y407V,T366I/K392M/T394W, F405A/Y407V, T366L/K392M/T394W, T366L/K392L/T394W,L351Y/Y407A, L351Y/Y407V, T366A/K409F, T366V/K409F, T366A/K409F,T350V/L351Y/F405A/Y407V and T350V/T366L/K392L/T394W, wherein residuenumbering is according to the EU index.

In other embodiments, the first Ig constant region or the fragment ofthe first Ig constant region and the second Ig constant region or thefragment of the second Ig constant region comprise the followingmutations

L235A_L235A_D265S_T350V_L351Y_F405A_Y407V in the first Ig constantregion and L235A_L235A_D265S_T350V_T366L_K392L_T394W in the second Igconstant region; or

L235A_L235A_D265S_T350V_T366L_K392L_T394W in the first Ig constantregion and L235A_L235A_D265S_T350V_L351Y_F405A_Y407V in the second Igconstant region.

Trispecific Antibodies

In some embodiments, provided herein are trispecific antibodies thatbind to CD79b, CD20, and CD3, and trispecific binding fragments thereof.This can be achieved by, for example, making a molecule which comprisesa first region binding specifically to CD79b, a second binding regionbinding specifically to CD3 and a third binding region bindingspecifically to the CD20. The antigen-binding regions can take any formthat allows specific recognition of the target, for example the bindingregion may be or may include a heavy chain variable domain, an Fv(combination of a heavy chain variable domain and a light chain variabledomain), an single-chain Fv (scFv), an Fab, a binding domain based on afibronectin type III domain (such as from fibronectin, or based on aconsensus of the type III domains from fibronectin, or from tenascin orbased on a consensus of the type III domains from tenascin, such as theCentyrin molecules from Janssen Biotech, Inc., see e.g. WO2010/051274and WO2010/093627). Accordingly, trispecific molecules comprising threedifferent antigen-binding regions which bind CD79b, CD20, and CD3,respectively, are provided.

In some embodiments, the CD79b×CD20×CD3-multispecific antibody comprisesa first heavy chain (HC1) and a light chain (LC) that pair to form afirst antigen-binding site that specifically binds a first antigen and asecond heavy chain (HC2) comprises a second antigen-binding site thatspecifically binds a second antigen. Either the HC1 or the HC2 mayfurther comprise a third antigen-binding site that specifically binds athird antigen. The HC1 and HC2 may each comprise a Fragmentcrystallizable (Fc) domain comprising a CH2-CH3 domain. In preferredembodiments, the CD79b×CD20×CD3-multispecific antibody is a trispecificantibody comprising a CD79b-specific arm comprising a first heavy chain(HC1) and a light chain (LC) that pair to form a first antigen-bindingsite that specifically binds CD79b, a second heavy chain (HC2) thatcomprises a second antigen-binding site that specifically binds a secondantigen, and the HC1 or the HC2 further comprises a thirdantigen-binding site that specifically binds a third antigen. In someembodiments, the second antigen is CD20, and the third antigen is CD3.In some embodiments, the second antigen is CD3, and the third antigen isCD20.

In some embodiment, the HC2 comprises the third antigen-binding sitethat specifically binds the third antigen. For example, the HC2 maycomprise, from N to C-terminus, the second antigen-binding site, the Fcdomain, a linker, and the third antigen-binding site.

In some embodiment, the HC1 comprises the third antigen-binding sitethat specifically binds the third antigen. For example, the HC1 maycomprise, from N to C-terminus, a heavy chain variable domain (VH)associated with the first antigen-binding site, a CH1 domain, the Fcdomain, a linker, and the third antigen-binding site.

In one embodiment, the CD79b×CD20×CD3-multispecific antibody is atrispecific antibody comprising a CD79b-specific arm comprising an HC1and a LC that pair to form a first antigen-binding site thatspecifically binds CD79b, an HC2 that comprises a second antigen-bindingsite that specifically binds CD3, and the HC2 further comprises a thirdantigen-binding site that specifically binds CD20.

In one embodiment, the CD79b×CD20×CD3-multispecific antibody is atrispecific antibody comprising a CD79b-specific arm comprising an HC1and a LC that pair to form a first antigen-binding site thatspecifically binds CD79b, an HC2 that comprises a second antigen-bindingsite that specifically binds CD20, and the HC2 further comprises a thirdantigen-binding site that specifically binds CD3.

In one embodiment, the CD79b×CD20×CD3-multispecific antibody is atrispecific antibody comprising a CD79b-specific arm comprising an HC1and a LC that pair to form a first antigen-binding site thatspecifically binds CD79b, an HC2 that comprises a second antigen-bindingsite that specifically binds CD20, and the HC1 further comprises a thirdantigen-binding site that specifically binds CD3.

In some embodiments, the first antigen-binding site comprises anantigen-binding fragment (Fab). In some embodiments, the secondantigen-binding site comprises a single-chain variable fragment (scFv).In some embodiments, the third antigen-binding site comprises asingle-chain variable fragment (scFv).

In one embodiment, the CD79b-binding arm comprises an antigen-bindingfragment (Fab), the CD3-binding arm comprises a single-chain variablefragment (scFv), and the CD20-binding arm comprises a single-chainvariable fragment (scFv).

Exemplary heavy chains and light chains for the exemplary tri-specificbinding proteins of the disclosure are shown in Table 31.

Generation of Multispecific Proteins that Comprise Antigen BindingFragments that Bind CD3ε.

The antigen binding fragments that bind CD3ε of the disclosure may beengineered into multispecific antibodies which are also encompassedwithin the scope of the invention.

The antigen binding fragments that bind CD3ε may be engineered into fulllength multispecific antibodies which are generated using Fab armexchange, in which substitutions are introduced into two monospecificbivalent antibodies within the Ig constant region CH3 domain whichpromote Fab arm exchange in vitro. In the methods, two monospecificbivalent antibodies are engineered to have certain substitutions at theCH3 domain that promote heterodimer stability; the antibodies areincubated together under reducing conditions sufficient to allow thecysteines in the hinge region to undergo disulfide bond isomerization;thereby generating the bispecific antibody by Fab arm exchange. Theincubation conditions may optimally be restored to non-reducing.Exemplary reducing agents that may be used are 2-mercaptoethylamine(2-MEA), dithiothreitol (DTT), dithioerythritol (DTE), glutathione,tris(2-carboxyethyl)phosphine (TCEP), L-cysteine andbeta-mercaptoethanol, preferably a reducing agent selected from thegroup consisting of: 2-mercaptoethylamine, dithiothreitol andtris(2-carboxyethyl)phosphine. For example, incubation for at least 90min at a temperature of at least 20° C. in the presence of at least 25mM 2-MEA or in the presence of at least 0.5 mM dithiothreitol at a pH offrom 5-8, for example at pH of 7.0 or at pH of 7.4 may be used.

CH3 mutations that may be used include technologies such as Knob-in-Holemutations (Genentech), electrostatically-matched mutations (Chugai,Amgen, NovoNordisk, Oncomed), the Strand Exchange Engineered Domain body(SEEDbody) (EMD Serono), Duobody® mutations (Genmab), and otherasymmetric mutations (e.g. Zymeworks).

Knob-in-hole mutations are disclosed for example in WO1996/027011 andinclude mutations on the interface of CH3 region in which an amino acidwith a small side chain (hole) is introduced into the first CH3 regionand an amino acid with a large side chain (knob) is introduced into thesecond CH3 region, resulting in preferential interaction between thefirst CH3 region and the second CH3 region. Exemplary CH3 regionmutations forming a knob and a hole are T366Y/F405A, T366W/F405W,F405W/Y407A, T394W/Y407T, T394S/Y407A, T366W/T394S, F405W/T394S and1366W/1366S_L368A_Y407V.

Heavy chain heterodimer formation may be promoted by using electrostaticinteractions by substituting positively charged residues on the firstCH3 region and negatively charged residues on the second CH3 region asdescribed in US2010/0015133, US2009/0182127, US2010/028637 orUS2011/0123532.

Other asymmetric mutations that can be used to promote heavy chainheterodimerization are L351Y_F405A_Y407V/T394W,T366I_K392M_T394W/F405A_Y407V, T366L_K392M_T394W/F405A_Y407V,L351Y_Y407A/T366A_K409F, L351Y_Y407A/T366V_K409F, Y407A/T366A_K409F, orT350V_L351Y_F405A_Y407V/T350V_T366L_K392L_T394W as described inUS2012/0149876 or US2013/0195849 (Zymeworks).

SEEDbody mutations involve substituting select IgG residues with IgAresidues to promote heavy chain heterodimerization as described inUS20070287170.

Other exemplary mutations that may be used are R409D_K370E/D399K_E357K,S354C_T366W/Y349C_T366S_L368A_Y407V,Y349C_T366W/S354C_T366S_L368A_Y407V, T366K/L351D, L351K/Y349E,L351K/Y349D, L351K/L368E, L351Y_Y407A/T366A_K409F,L351Y_Y407A/T366V_K409F, K392D/D399K, K392D/E356K,K253E_D282K_K322D/D239K_E240K_K292D, K392D_K409D/D356K_D399K asdescribed in WO2007/147901, WO 2011/143545, WO2013157954, WO2013096291and US2018/0118849.

Duobody® mutations (Genmab) are disclosed for example in U.S. Pat. No.9,150,663 and US2014/0303356 and include mutations F405L/K409R,wild-type/F405L_R409K, T350I_K370T_F405L/K409R, K370W/K409R,D399AFGHILMNRSTVWY/K409R, T366ADEFGHILMQVY/K409R,L368ADEGHNRSTVQ/K409AGRH, D399FHKRQ/K409AGRH, F405IKLSTVW/K409AGRH andY407LWQ/K409AGRH.

Additional bispecific or multispecific structures into which the antigenbinding domains that bind

CD3ε can be incorporated include Dual Variable Domain Immunoglobulins(DVD) (Int. Pat. Publ. No. WO2009/134776; DVDs are full lengthantibodies comprising the heavy chain having a structureVH1-linker-VH2-CH and the light chain having the structureVL1-linker-VL2-CL; linker being optional), structures that includevarious dimerization domains to connect the two antibody arms withdifferent specificity, such as leucine zipper or collagen dimerizationdomains (Int. Pat. Publ. No. WO2012/022811, U.S. Pat. Nos. 5,932,448;6,833,441), two or more domain antibodies (dAbs) conjugated together,diabodies, heavy chain only antibodies such as camelid antibodies andengineered camelid antibodies, Dual Targeting (DT)-Ig (GSK/Domantis),Two-in-one Antibody (Genentech), Cross-linked Mabs (Karmanos CancerCenter), mAb2 (F-Star) and CovX-body (CovX/Pfizer), IgG-like Bispecific(InnClone/Eli Lilly), Ts2Ab (MedImmune/AZ) and BsAb (Zymogenetics),HERCULES (Biogen Idec) and TvAb (Roche), ScFv/Fc Fusions (AcademicInstitution), SCORPION (Emergent BioSolutions/Trubion,Zymogenetics/BMS), Dual Affinity Retargeting Technology (Fc-DART)(MacroGenics) and Dual(ScFv)2-Fab (National Research Center for AntibodyMedicine—China), Dual-Action or Bis-Fab (Genentech), Dock-and-Lock (DNL)(ImmunoMedics), Bivalent Bispecific (Biotecnol) and Fab-Fv(UCB-Celltech). ScFv-, diabody-based, and domain antibodies, include butare not limited to, Bispecific T Cell Engager (BiTE) (Micromet), TandemDiabody (Tandab) (Affimed), Dual Affinity Retargeting Technology (DART)(MacroGenics), Single-chain Diabody (Academic), TCR-like Antibodies(AIT, ReceptorLogics), Human Serum Albumin ScFv Fusion (Merrimack) andCOMBODY (Epigen Biotech), dual targeting nanobodies (Ablynx), dualtargeting heavy chain only domain antibodies.

The antigen binding domains that bind CD3ε of the disclosure may also beengineered into multispecific proteins which comprise three polypeptidechains. In such designs, at least one antigen binding domain is in theform of a scFv. Exemplary designs include (in which “1” indicates thefirst antigen binding domain, “2” indicates the second antigen bindingdomain and “3” indicates the third antigen binding domain:

Design 1: Chain A) scFv1-CH2-CH3; Chain B) VL2-CL; Chain C)VH2-CH1-hinge-CH2-CH3

Design 2: Chain A) scFv1-hinge-CH2-CH3; Chain B) VL2-CL; Chain C)VH2-CH1-hinge-CH2-CH3

Design 3: Chain A) scFv1-CH1-hinge-CH2-CH3; Chain B) VL2-CL; Chain C)VH2-CH1-hinge-CH2-CH3

Design 4: Chain A) CH2-CH3-scFv1; Chain B) VL2-CL; Chain C)VH2-CH1-hinge-CH2-CH3

CH3 engineering may be incorporated to the Designs 1-4, such asmutations L351Y_F405A_Y407V/T394W, T366I_K392M_T394W/F405A_Y407V,T366L_K392M_T394W/F405A_Y407V, L351Y_Y407A/T366A_K409F,L351Y_Y407A/T366V_K409F, Y407A/T366A_K409F, orT350V_L351Y_F405A_Y407V/T350V_T366L_K392L_T394W as described inUS2012/0149876 or US2013/0195849 (Zymeworks).

Isotypes, Allotypes and Fc Engineering

The Ig constant region or the fragment of the Ig constant region, suchas the Fc region present in the proteins of the disclosure may be of anyallotype or isotype.

In other embodiments, the Ig constant region or the fragment of the Igconstant region is an IgG1 isotype.

In other embodiments, the Ig constant region or the fragment of the Igconstant region is an IgG2 isotype.

In other embodiments, the Ig constant region or the fragment of the Igconstant region is an IgG3 isotype.

In other embodiments, the Ig constant region or the fragment of the Igconstant region is an IgG4 isotype.

The Ig constant region or the fragment of the Ig constant region may beof any allotype. It is expected that allotype has no influence onproperties of the Ig constant region, such as binding or Fc-mediatedeffector functions. Immunogenicity of therapeutic proteins comprising Igconstant regions of fragments thereof is associated with increased riskof infusion reactions and decreased duration of therapeutic response(Baert et al., (2003) N Engl J Med 348:602-08). The extent to whichtherapeutic proteins comprising Ig constant regions of fragments thereofinduce an immune response in the host may be determined in part by theallotype of the Ig constant region (Stickler et al., (2011) Genes andImmunity 12:213-21). Ig constant region allotype is related to aminoacid sequence variations at specific locations in the constant regionsequences of the antibody. Table 3 shows select IgG1, IgG2 and IgG4allotypes.

TABLE 3 Amino acid residue at position of diversity (residue numbering:EU Index) IgG2 IgG4 IgG1 Allotype 189 282 309 422 214 356 358 431 G2m(n)T M G2m(n−) P V G2m(n)/(n−) T V nG4m(a) L R G1m(17) K E M A G1m(17, 1) KD L A G1m(3) R E M A

C-terminal lysine (CTL) may be removed from the Ig constant region byendogenous circulating carboxypeptidases in the blood stream (Cai etal., (2011) Biotechnol Bioeng 108:404-412). During manufacturing, CTLremoval may be controlled to less than the maximum level by control ofconcentration of extracellular Zn²⁺, EDTA or EDTA-Fe³⁺ as described inU.S. Patent Publ. No. US20140273092. CTL content of proteins may bemeasured using known methods.

In other embodiments, the antigen binding fragment that binds CD3εconjugated to the Ig constant region has a C-terminal lysine contentfrom about 10% to about 90%. In other embodiments, the C-terminal lysinecontent is from about 20% to about 80%. In other embodiments, theC-terminal lysine content is from about 40% to about 70%. In otherembodiments, the C-terminal lysine content is from about 55% to about70%. In other embodiments, the C-terminal lysine content is about 60%.

Fc region mutations may be made to the antigen binding domains that bindCD3ε conjugated to the Ig constant region or to the fragment of the Igconstant region to modulate their effector functions such as ADCC, ADCPand/or ADCP and/or pharmacokinetic properties. This may be achieved byintroducing mutation(s) into the Fc that modulate binding of the mutatedFc to activating FcγRs (FcγRI, FcγRIIa, FcγRIII), inhibitory FcγRIIband/or to FcRn.

In other embodiments, the antigen binding domain that binds CD3εconjugated to the Ig constant region or the fragment of the Ig constantregion comprises at least one mutation in the Ig constant region or inthe fragment of the Ig constant region.

In other embodiments, the at least one mutation is in the Fc region.

In other embodiments, the antigen binding domain that binds CD3εconjugated to the Ig constant region or to the fragment of the Igconstant region comprises at least one, two, three, four, five, six,seven, eight, nine, ten, eleven, twelve, thirteen, fourteen or fifteenmutations in the Fc region.

In other embodiments, the antigen binding domain that binds CD3εconjugated to the Ig constant region or to the fragment of the Igconstant region comprises at least one mutation in the Fc region thatmodulates binding of the antibody to FcRn.

Fc positions that may be mutated to modulate half-life (e.g. binding toFcRn) include positions 250, 252, 253, 254, 256, 257, 307, 376, 380,428, 434 and 435. Exemplary mutations that may be made singularly or incombination are mutations T250Q, M252Y, I253A, S254T, T256E, P257I,T307A, D376V, E380A, M428L, H433K, N434S, N434A, N434H, N434F, H435A andH435R. Exemplary singular or combination mutations that may be made toincrease the half-life are mutations M428L/N434S, M252Y/S254T/T256E,T250Q/M428L, N434A and T307A/E380A/N434A. Exemplary singular orcombination mutations that may be made to reduce the half-life aremutations H435A, P257I/N434H, D376V/N434H,M252Y/S254T/T256E/H433K/N434F, T308P/N434A and H435R.

In other embodiments, the antigen binding domain that binds CD3εconjugated to the Ig constant region or to the fragment of the Igconstant region comprises M252Y/S254T/T256E mutation.

In other embodiments, the antigen binding domain that binds CD3εconjugated to the Ig constant region or to the fragment of the Igconstant region comprises at least one mutation in the Fc region thatreduces binding of the protein to an activating Fcγ receptor (FcγR)and/or reduces Fc effector functions such as C1q binding, complementdependent cytotoxicity (CDC), antibody-dependent cell-mediatedcytotoxicity (ADCC) or phagocytosis (ADCP).

Fc positions that may be mutated to reduce binding of the protein to theactivating FcγR and subsequently to reduce effector function includepositions 214, 233, 234, 235, 236, 237, 238, 265, 267, 268, 270, 295,297, 309, 327, 328, 329, 330, 331 and 365. Exemplary mutations that maybe made singularly or in combination are mutations K214T, E233P, L234V,L234A, deletion of G236, V234A, F234A, L235A, G237A, P238A, P238S,D265A, S267E, H268A, H268Q, Q268A, N297A, A327Q, P329A, D270A, Q295A,V309L, A327S, L328F, A330S and P331S in IgG1, IgG2, IgG3 or IgG4.Exemplary combination mutations that result in proteins with reducedADCC are mutations L234A/L235A on IgG1, L234A/L235A/D265S on IgG1,V234A/G237A/P238S/H268A/V309L/A330S/P331S on IgG2, F234A/L235A on IgG4,S228P/F234A/L235A on IgG4, N297A on all Ig isotypes, V234A/G237A onIgG2, K214T/E233P/L234V/L235A/G236-deleted/A327G/P331A/D365E/L358M onIgG1, H268Q/V309L/A330S/P331S on IgG2, S267E/L328F on IgG1,L234F/L235E/D265A on IgG1, L234A/L235A/G237A/P238S/H268A/A330S/P331S onIgG1, S228P/F234A/L235A/G237A/P238S on IgG4, andS228P/F234A/L235A/G236-deleted/G237A/P238S on IgG4. Hybrid IgG2/4 Fcdomains may also be used, such as Fc with residues 117-260 from IgG2 andresidues 261-447 from IgG4.

Exemplary mutation that result in proteins with reduced CDC is a K322Amutation.

Well-known S228P mutation may be made in IgG4 to enhance IgG4 stability.

In other embodiments, the antigen binding domain that binds CD3εconjugated to the Ig constant region or to the fragment of the Igconstant region comprises at least one mutation selected from the groupconsisting of K214T, E233P, L234V, L234A, deletion of G236, V234A,F234A, L235A, G237A, P238A, P238S, D265A, S267E, H268A, H268Q, Q268A,N297A, A327Q, P329A, D270A, Q295A, V309L, A327S, L328F, K322, A330S andP331S.

In other embodiments, the antigen binding domain that binds CD3εconjugated to the Ig constant region or to the fragment of the Igconstant region comprises L234A/L235A/D265S mutation.

In other embodiments, the antigen binding domain that binds CD3εconjugated to the Ig constant region or to the fragment of the Igconstant region comprises L234A/L235A mutation.

In other embodiments, the antigen binding domain that binds CD3εconjugated to the Ig constant region or to the fragment of the Igconstant region comprises at least one mutation in the Fc region thatenhances binding of the protein to an Fcγ receptor (FcγR) and/orenhances Fc effector functions such as C1q binding, complement dependentcytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC)and/or phagocytosis (ADCP).

Fc positions that may be mutated to increase binding of the protein tothe activating FcγR and/or enhance Fc effector functions includepositions 236, 239, 243, 256, 290, 292, 298, 300, 305, 312, 326, 330,332, 333, 334, 345, 360, 339, 378, 396 or 430 (residue numberingaccording to the EU index). Exemplary mutations that may be madesingularly or in combination are G236A, S239D, F243L, T256A, K290A,R292P, S298A, Y300L, V305L, K326A, A330K, I332E, E333A, K334A, A339T andP396L. Exemplary combination mutations that result in proteins withincreased ADCC or ADCP are a S239D/I332E, S298A/E333A/K334A,F243L/R292P/Y300L, F243L/R292P/Y300L/P396L,F243L/R292P/Y300L/V305I/P396L and G236A/S239D/I332E.

Fc positions that may be mutated to enhance CDC include positions 267,268, 324, 326, 333, 345 and 430. Exemplary mutations that may be madesingularly or in combination are S267E, F1268F, S324T, K326A, K326W,E333A, E345K, E345Q, E345R, E345Y, E430S, E430F and E430T. Exemplarycombination mutations that result in proteins with increased CDC areK326A/E333A, K326W/E333A, H268F/S324T, S267E/H268F, S267E/S324T andS267E/H268F/S324T.

The specific mutations described herein are mutations when compared tothe IgG1, IgG2 and IgG4 wild-type amino acid sequences of SEQ ID NOs:237, 238, and 239, respectively.

wild-type IgG1, SEQ ID NO: 237ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK wild-type IgG2; SEQ ID NO: 238ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDISVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK wild-type IgG4; SEQ ID NO: 239ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

Binding of the antibody to FcγR or FcRn may be assessed on cellsengineered to express each receptor using flow cytometry. In anexemplary binding assay, 2×10⁵ cells per well are seeded in 96-wellplate and blocked in BSA Stain Buffer (BD Biosciences, San Jose, USA)for 30 min at 4° C. Cells are incubated with a test antibody on ice for1.5 hour at 4° C. After being washed twice with BSA stain buffer, thecells are incubated with R-PE labeled anti-human IgG secondary antibody(Jackson Immunoresearch Laboratories) for 45 min at 4° C. The cells arewashed twice in stain buffer and then resuspended in 150 μL of StainBuffer containing 1:200 diluted DRAQ7 live/dead stain (Cell SignalingTechnology, Danvers, USA). PE and DRAQ7 signals of the stained cells aredetected by Miltenyi MACSQuant flow cytometer (Miltenyi Biotec, Auburn,USA) using B2 and B4 channel respectively. Live cells are gated on DRAQ7exclusion and the geometric mean fluorescence signals are determined forat least 10,000 live events collected. FlowJo software (Tree Star) isused for analysis. Data is plotted as the logarithm of antibodyconcentration versus mean fluorescence signals. Nonlinear regressionanalysis is performed.

Glycoengineering

The ability of the antigen binding domain that binds CD3ε conjugated tothe Ig constant region or to the fragment of the Ig constant region tomediate ADCC can be enhanced by engineering the Ig constant region orthe fragment of the Ig constant region oligosaccharide component. HumanIgG1 or IgG3 are N-glycosylated at Asn297 with the majority of theglycans in the well-known biantennary G0, G0F, G1, G1F, G2 or G2F forms.Ig constant region containing proteins may be produced by non-engineeredCHO cells typically have a glycan fucose content of about at least 85%.The removal of the core fucose from the biantennary complex-typeoligosaccharides attached to the antigen binding domain that binds CD3εconjugated to the Ig constant region or to the fragment of the Igconstant region enhances the ADCC of the protein via improved FcγRIIIabinding without altering antigen binding or CDC activity. Such proteinscan be achieved using different methods reported to lead to thesuccessful expression of relatively high defucosylated immunoglobulinsbearing the biantennary complex-type of Fc oligosaccharides such ascontrol of culture osmolality (Konno et al., Cytotechnology 64(:249-65,2012), application of a variant CHO line Lec13 as the host cell line(Shields et al., J Biol Chem 277:26733-26740, 2002), application of avariant CHO line EB66 as the host cell line (Olivier et al., MAbs; 2(4):405-415, 2010; PMID:20562582), application of a rat hybridoma cell lineYB2/0 as the host cell line (Shinkawa et al., J Biol Chem 278:3466-3473,2003), introduction of small interfering RNA specifically against the a1,6-fucosyltrasferase (FUT8) gene (Mori et al., Biotechnol Bioeng88:901-908, 2004), or coexpression ofβ-1,4-N-acetylglucosaminyltransferase III and Golgi α-mannosidase II ora potent alpha-mannosidase I inhibitor, kifunensine (Ferrara et al., JBiol Chem 281:5032-5036, 2006, Ferrara et al., Biotechnol Bioeng93:851-861, 2006; Xhou et al., Biotechnol Bioeng 99:652-65, 2008).

In other embodiments, the antigen binding domain that binds CD3εconjugated to the Ig constant region or to the fragment of the Igconstant region of the disclosure has a biantennary glycan structurewith fucose content of about between 1% to about 15%, for example about15%, 14%, 13%, 12%, 11% 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1%. Inother embodiments, the antigen binding domain that binds CD3ε conjugatedto the Ig constant region or to the fragment of the Ig constant regionhas a glycan structure with fucose content of about 50%, 40%, 45%, 40%,35%, 30%, 25%, or 20%.

“Fucose content” means the amount of the fucose monosaccharide withinthe sugar chain at Asn297. The relative amount of fucose is thepercentage of fucose-containing structures related to allglycostructures. These may be characterized and quantified by multiplemethods, for example: 1) using MALDI-TOF of N-glycosidase F treatedsample (e.g. complex, hybrid and oligo- and high-mannose structures) asdescribed in Int Pat. Publ. No. WO2008/077546 2); 2) by enzymaticrelease of the Asn297 glycans with subsequent derivatization anddetection/quantitation by HPLC (UPLC) with fluorescence detection and/orHPLC-MS (UPLC-MS); 3) intact protein analysis of the native or reducedmAb, with or without treatment of the Asn297 glycans with Endo S orother enzyme that cleaves between the first and the second GlcNAcmonosaccharides, leaving the fucose attached to the first GlcNAc; 4)digestion of the mAb to constituent peptides by enzymatic digestion(e.g., trypsin or endopeptidase Lys-C), and subsequent separation,detection and quantitation by HPLC-MS (UPLC-MS); 5) Separation of themAb oligosaccharides from the mAb protein by specific enzymaticdeglycosylation with PNGase F at Asn 297. The oligosaccharides thusreleased can be labeled with a fluorophore, separated and identified byvarious complementary techniques which allow: fine characterization ofthe glycan structures by matrix-assisted laser desorption ionization(MALDI) mass spectrometry by comparison of the experimental masses withthe theoretical masses, determination of the degree of sialylation byion exchange HPLC (GlycoSep C), separation and quantification of theoligosaccharide forms according to hydrophilicity criteria bynormal-phase HPLC (GlycoSep N), and separation and quantification of theoligosaccharides by high performance capillary electrophoresis-laserinduced fluorescence (HPCE-LIF).

“Low fucose” or “low fucose content” as used herein refers to theantigen binding domain that bind CD3ε conjugated to the Ig constantregion or to the fragment of the Ig constant region with fucose contentof about between 1%-15%.

“Normal fucose” or ‘normal fucose content” as used herein refers to theantigen binding domain that bind CD3ε conjugated to the Ig constantregion or to the fragment of the Ig constant region with fucose contentof about over 50%, typically about over 80% or over 85%.

Anti-Idiotypic Antibodies

Anti-idiotypic antibodies are antibodies that specifically bind to theantigen binding domain that binds CD3ε of the disclosure.

The invention also provides an anti-idiotypic antibody that specificallybinds to the antigen binding domain that binds CD3ε of the disclosure.

The invention also provides an anti-idiotypic antibody that specificallybinds to the antigen binding domain that binds CD3ε comprising

-   -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59;    -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 56;    -   the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 88 and the VL of SEQ ID NO: 58; or    -   the VH of SEQ ID NO: 242 and the VL of SEQ ID NO: 58.

An anti-idiotypic (Id) antibody is an antibody which recognizes theantigenic determinants (e.g. the paratope or CDRs) of the antibody. TheId antibody may be antigen-blocking or non-blocking. Theantigen-blocking Id may be used to detect the free antigen bindingdomain in a sample (e.g. the antigen binding domain that binds CD3ε ofthe disclosure). The non-blocking Id may be used to detect the totalantibody (free, partially bond to antigen, or fully bound to antigen) ina sample. An Id antibody may be prepared by immunizing an animal withthe antibody to which an anti-Id is being prepared.

An anti-Id antibody may also be used as an immunogen to induce an immuneresponse in yet another animal, producing a so-called anti-anti-Idantibody. An anti-anti-Id may be epitopically identical to the originalantigen binding domain which induced the anti-Id. Thus, by usingantibodies to the idiotypic determinants of the antigen binding domain,it is possible to identify other clones expressing antigen bindingdomains of identical specificity. Anti-Id antibodies may be varied(thereby producing anti-Id antibody variants) and/or derivatized by anysuitable technique, such as those described elsewhere herein.

Immunoconjugates

The antigen binding domains that bind CD3ε of the disclosure, theproteins comprising the antigen binding domains that bind CD3ε or themultispecific proteins that comprise the antigen binding domains thatbind CD3ε (collectively referred herein as to CD3ε binding proteins) maybe conjugated to a heterologous molecule.

In other embodiments, the heterologous molecule is a detectable label ora cytotoxic agent.

The invention also provides an antigen binding domain that binds CD3εconjugated to a detectable label.

The invention also provides a protein comprising an antigen bindingdomain that binds CD3ε conjugated to a detectable label.

The invention also provides a multispecific protein comprising anantigen binding domain that binds CD3ε conjugated to a detectable label.

The invention also provides an antigen binding domain that binds CD3εconjugated to a cytotoxic agent.

The invention also provides a protein comprising an antigen bindingdomain that binds CD3ε conjugated to a cytotoxic agent.

The invention also provides a multispecific protein comprising anantigen binding domain that binds CD3ε conjugated to a cytotoxic agent.

CD3ε binding proteins of the disclosure may be used to directtherapeutics to tumor antigen expressing cells. Alternatively, CD3εexpressing cells may be targeted with a CD3ε binding protein of thedisclosure coupled to a therapeutic intended to modify cell functiononce internalized.

In other embodiments, the detectable label is also a cytotoxic agent.

The CD3ε binding proteins of the disclosure conjugated to a detectablelabel may be used to evaluate expression of CD3ε on a variety ofsamples.

Detectable label includes compositions that when conjugated to the CD3εbinding proteins of the disclosure renders the latter detectable, viaspectroscopic, photochemical, biochemical, immunochemical, or chemicalmeans.

Exemplary detectable labels include radioactive isotopes, magneticbeads, metallic beads, colloidal particles, fluorescent dyes,electron-dense reagents, enzymes (for example, as commonly used in anELISA), biotin, digoxigenin, haptens, luminescent molecules,chemiluminescent molecules, fluorochromes, fluorophores, fluorescentquenching agents, colored molecules, radioactive isotopes, scintillates,avidin, streptavidin, protein A, protein G, antibodies or fragmentsthereof, polyhistidine, Ni²⁺, Flag tags, myc tags, heavy metals,enzymes, alkaline phosphatase, peroxidase, luciferase, electrondonors/acceptors, acridinium esters, and colorimetric substrates.

A detectable label may emit a signal spontaneously, such as when thedetectable label is a radioactive isotope. In other cases, thedetectable label emits a signal as a result of being stimulated by anexternal field.

Exemplary radioactive isotopes may be γ-emitting, Auger-emitting,β-emitting, an alpha-emitting or positron-emitting radioactive isotope.Exemplary radioactive isotopes include ³H, ¹¹C, ¹³C, ¹⁵N, ¹⁸F, ¹⁹F,⁵⁵Co, ⁵⁷Co, ⁶⁰Co, ⁶¹Cu, ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, ⁶⁸Ga, ⁷²As, ⁷⁵Br, ⁸⁶Y, ⁸⁹Zr,⁹⁰Sr, ^(94m)Tc, ^(99m)Tc, ¹¹⁵In, ¹²³I, ¹²⁴I, ¹²⁵I, ¹³¹I, ²¹¹At, ²¹²Bi,²¹³Bi, ²²³Ra, ²²⁶Ra, ²²⁵Ac and ²²⁷Ac.

Exemplary metal atoms are metals with an atomic number greater than 20,such as calcium atoms, scandium atoms, titanium atoms, vanadium atoms,chromium atoms, manganese atoms, iron atoms, cobalt atoms, nickel atoms,copper atoms, zinc atoms, gallium atoms, germanium atoms, arsenic atoms,selenium atoms, bromine atoms, krypton atoms, rubidium atoms, strontiumatoms, yttrium atoms, zirconium atoms, niobium atoms, molybdenum atoms,technetium atoms, ruthenium atoms, rhodium atoms, palladium atoms,silver atoms, cadmium atoms, indium atoms, tin atoms, antimony atoms,tellurium atoms, iodine atoms, xenon atoms, cesium atoms, barium atoms,lanthanum atoms, hafnium atoms, tantalum atoms, tungsten atoms, rheniumatoms, osmium atoms, iridium atoms, platinum atoms, gold atoms, mercuryatoms, thallium atoms, lead atoms, bismuth atoms, francium atoms, radiumatoms, actinium atoms, cerium atoms, praseodymium atoms, neodymiumatoms, promethium atoms, samarium atoms, europium atoms, gadoliniumatoms, terbium atoms, dysprosium atoms, holmium atoms, erbium atoms,thulium atoms, ytterbium atoms, lutetium atoms, thorium atoms,protactinium atoms, uranium atoms, neptunium atoms, plutonium atoms,americium atoms, curium atoms, berkelium atoms, californium atoms,einsteinium atoms, fermium atoms, mendelevium atoms, nobelium atoms, orlawrencium atoms.

In other embodiments, the metal atoms may be alkaline earth metals withan atomic number greater than twenty.

In other embodiments, the metal atoms may be lanthanides.

In other embodiments, the metal atoms may be actinides.

In other embodiments, the metal atoms may be transition metals.

In other embodiments, the metal atoms may be poor metals.

In other embodiments, the metal atoms may be gold atoms, bismuth atoms,tantalum atoms, and gadolinium atoms.

In other embodiments, the metal atoms may be metals with an atomicnumber of 53 (i.e. iodine) to 83 (i.e. bismuth).

In other embodiments, the metal atoms may be atoms suitable for magneticresonance imaging.

The metal atoms may be metal ions in the form of +1, +2, or +3 oxidationstates, such as Ba²⁺, Bi³⁺, Cs⁺, Ca²⁺, Cr²⁺, Cr³⁺, Cr⁶⁺, Co²⁺, Co³⁺,Cu⁺, Cu²⁺, Cu³⁺, Ga³⁺, Gd³⁺, Au⁺, Au³⁺, Fe²⁺, Fe³⁺, F³⁺, Pb²⁺, Mn²⁺,Mn³⁺, Mn⁴⁺, Mn⁷⁺, Hg²⁺, Ni²⁺, Ni³⁺, Ag⁺, Sn²⁺, Sn⁴⁺, and Zn²⁺. The metalatoms may comprise a metal oxide, such as iron oxide, manganese oxide,or gadolinium oxide.

Suitable dyes include any commercially available dyes such as, forexample, 5(6)-carboxyfluorescein, IRDye 680RD maleimide or IRDye 800CW,ruthenium polypyridyl dyes, and the like.

Suitable fluorophores are fluorescein isothiocyanate (FITC), fluoresceinthiosemicarbazide, rhodamine, Texas Red, CyDyes (e.g., Cy3, Cy5, Cy5.5),Alexa Fluors (e.g., Alexa488, Alexa555, Alexa594; Alexa647), nearinfrared (NIR) (700-900 nm) fluorescent dyes, and carbocyanine andaminostyryl dyes.

The antigen binding domain that binds CD3ε conjugated to a detectablelabel may be used as an imaging agent.

The protein comprising an antigen binding domain that binds CD3εconjugated to a detectable label may be used as an imaging agent.

The multispecific protein comprising an antigen binding domain thatbinds CD3ε conjugated to a detectable label may be used as an imagingagent.

In other embodiments, the cytotoxic agent is a chemotherapeutic agent, adrug, a growth inhibitory agent, a toxin (e.g., an enzymatically activetoxin of bacterial, fungal, plant, or animal origin, or fragmentsthereof), or a radioactive isotope (i.e., a radioconjugate).

In other embodiments, the cytotoxic agent is daunomycin, doxorubicin,methotrexate, vindesine, bacterial toxins such as diphtheria toxin,ricin, geldanamycin, maytansinoids or calicheamicin. The cytotoxic agentmay elicit their cytotoxic and cytostatic effects by mechanismsincluding tubulin binding, DNA binding, or topoisomerase inhibition.

In other embodiments, the cytotoxic agent is an enzymatically activetoxin such as diphtheria A chain, nonbinding active fragments ofdiphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricinA chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordiiproteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII,and PAP-S), Momordica charantia inhibitor, curcin, crotin, Sapaonariaofficinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin,enomycin, and the tricothecenes.

In other embodiments, the cytotoxic agent is a radionuclide, such as²¹²Bi, ¹³¹I, ¹³¹In, ⁹⁰Y, and ¹⁸⁶Re.

In other embodiments, the cytotoxic agent is dolastatins or dolostatinpeptidic analogs and derivatives, auristatin or monomethyl auristatinphenylalanine Exemplary molecules are disclosed in U.S. Pat. Nos.5,635,483 and 5,780,588. Dolastatins and auristatins have been shown tointerfere with microtubule dynamics, GTP hydrolysis, and nuclear andcellular division (Woyke et al (2001) Antimicrob Agents and Chemother.45(12):3580-3584) and have anticancer and antifungal activity. Thedolastatin or auristatin drug moiety may be attached to the antibody ofthe invention through the N (amino) terminus or the C (carboxyl)terminus of the peptidic drug moiety (WO02/088172), or via any cysteineengineered into the antibody.

The CD3ε binding proteins of the disclosure may be conjugated to adetectable label using known methods.

In other embodiments, the detectable label is complexed with a chelatingagent.

In other embodiments, the detectable label is conjugated to the CD3εbinding proteins of the disclosure via a linker.

The detectable label or the cytotoxic moiety may be linked directly, orindirectly, to the CD3ε binding proteins of the disclosure using knownmethods. Suitable linkers are known in the art and include, for example,prosthetic groups, non-phenolic linkers (derivatives ofN-succimidyl-benzoates; dodecaborate), chelating moieties of bothmacrocyclics and acyclic chelators, such as derivatives of1,4,7,10-tetraazacyclododecane-1,4,7,10,tetraacetic acid (DOTA),derivatives of diethylenetriaminepentaacetic avid (DTPA), derivatives ofS-2-(4-Isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triaceticacid (NOTA) and derivatives of1,4,8,11-tetraazacyclodocedan-1,4,8,11-tetraacetic acid (TETA),N-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane(IT), bifunctional derivatives of imidoesters (such as dimethyladipimidate HCl), active esters (such as disuccinimidyl suberate),aldehydes (such as glutaraldehyde), bis-azido compounds (such asbis(p-azidobenzoyl)hexanediamine), bis-diazonium derivatives (such asbis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such astoluene 2,6-diisocyanate), and bis-active fluorine compounds (such as1,5-difluoro-2,4-dinitrobenzene) and other chelating moieties. Suitablepeptide linkers are well known.

In other embodiments, the CD3ε binding proteins of the disclosure isremoved from the blood via renal clearance.

Kits

The invention also provides a kit comprising the antigen binding domainthat binds CD3ε.

The invention also provides a kit comprising the protein comprising anantigen binding domain that binds CD3ε.

The invention also provides a kit comprising the multispecific proteincomprising an antigen binding domain that binds CD3ε.

The kit may be used for therapeutic uses and as diagnostic kits.

The kit may be used to detect the presence of CD3ε in a sample.

In other embodiments, the kit comprises the CD3ε binding protein of thedisclosure and reagents for detecting the CD3ε binding protein. The kitcan include one or more other elements including: instructions for use;other reagents, e.g., a label, a therapeutic agent, or an agent usefulfor chelating, or otherwise coupling, an antibody to a label ortherapeutic agent, or a radioprotective composition; devices or othermaterials for preparing the antibody for administration;pharmaceutically acceptable carriers; and devices or other materials foradministration to a subject.

In other embodiments, the kit comprises the antigen binding domain thatbinds CD3ε in a container and instructions for use of the kit.

In other embodiments, the kit comprises the protein comprising anantigen binding domain that binds CD3ε in a container and instructionsfor use of the kit.

In other embodiments, the kit comprises the multispecific proteincomprising an antigen binding domain that binds CD3ε in a container andinstructions for use of the kit.

In other embodiments, the antigen binding domain that binds CD3ε in thekit is labeled.

In other embodiments, the protein comprising an antigen binding domainthat binds CD3ε in the kit is labeled.

In other embodiments, the multispecific protein comprising an antigenbinding domain that binds CD3ε in the kit is labeled.

In other embodiments, the kit comprises the antigen binding domain thatbinds CD3ε comprising

-   -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59;    -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 56;    -   the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 88 and the VL of SEQ ID NO: 58; or    -   the VH of SEQ ID NO: 242 and the VL of SEQ ID NO: 58.

In other embodiments, the kit comprises the antigen binding domain thatbinds CD3ε comprising SEQ ID NOs: 96, 97, 98, 99, 100, 101, 102, 103,104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117,118, 119, 120, 121, 122, 123, 124, 125, or 126.

Methods of Detecting CD3ε

The invention also provides a method of detecting CD3ε in a sample,comprising obtaining the sample, contacting the sample with the antigenbinding domain that binds CD3ε of the disclosure and detecting the boundCD3ε in the sample.

In other embodiments, the sample may be derived from urine, blood,serum, plasma, saliva, ascites, circulating cells, synovial fluid,circulating cells, cells that are not tissue associated (i.e., freecells), tissues (e.g., surgically resected tissue, biopsies, includingfine needle aspiration), histological preparations, and the like.

The antigen binding domain that binds CD3ε of the disclosure may bedetected using known methods. Exemplary methods include direct labelingof the antibodies using fluorescent or chemiluminescent labels, orradiolabels, or attaching to the antibodies of the invention a moietywhich is readily detectable, such as biotin, enzymes or epitope tags.Exemplary labels and moieties are ruthenium, ¹¹¹In-DOTA,¹¹¹In-diethylenetriaminepentaacetic acid (DTPA), horseradish peroxidase,alkaline phosphatase and beta-galactosidase, poly-histidine (HIS tag),acridine dyes, cyanine dyes, fluorone dyes, oxazin dyes, phenanthridinedyes, rhodamine dyes and Alexafluor® dyes.

The antigen binding domain that binds CD3ε of the disclosure may be usedin a variety of assays to detect CD3ε in the sample. Exemplary assaysare western blot analysis, radioimmunoassay, surface plasmon resonance,immunoprecipitation, equilibrium dialysis, immunodiffusion,electrochemiluminescence (ECL) immunoassay, immunohistochemistry,fluorescence-activated cell sorting (FACS) or ELISA assay.

Polynucleotides, Vectors, Host Cells

The disclosure also provides an isolated polynucleotide encoding any ofthe CD3ε binding proteins of the disclosure. The CD3ε binding proteinincludes the antigen binding domains that bind CD3ε, the proteinscomprising the antigen binding domains that bind CD3ε, the multispecificproteins that comprise the antigen binding domains that bind CD3ε of thedisclosure.

The invention also provides an isolated polynucleotide encoding any ofCD3ε biding proteins or fragments thereof.

The invention also provides an isolated polynucleotide encoding the VHof SEQ ID NOs: 55, 54, or 48.

The invention also provides an isolated polynucleotide encoding the VLof SEQ ID NOs: 59, 58 or 56.

The invention also provides an isolated polynucleotide encoding the VHof SEQ ID NO: 55.

The invention also provides an isolated polynucleotide encoding the VHof SEQ ID NO: 54.

The invention also provides an isolated polynucleotide encoding the VHof SEQ ID NO: 48.

The invention also provides an isolated polynucleotide encoding the VLof SEQ ID NO: 59.

The invention also provides an isolated polynucleotide encoding the VLof SEQ ID NO: 58.

The invention also provides an isolated polynucleotide encoding the VLof SEQ ID NO: 56.

The invention also provides an isolated polynucleotide encoding the VHof SEQ ID NOs: 55, 54, or 48 and the VL of SEQ ID NOs: 24, 27, 28, 29 or30.

The invention also provides for an isolated polynucleotide encoding

-   -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59;    -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 56;    -   the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 88 and the VL of SEQ ID NO: 58; or    -   the VH of SEQ ID NO: 242 and the VL of SEQ ID NO: 58.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NOs: 96, 97, 98, 99, 100, 101, 102, 103, 104, 105,106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119,120, 121, 122, 123, 124, 125, or 126.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 96.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 97.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 98.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 99.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 100.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 101.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 102.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 103.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 104.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 105.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 106.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 107.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 108.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 109.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 110.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 112.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 113.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 114.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 115.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 116.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 117.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 118.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 119.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 120.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 121.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 122.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 123.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 124.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 125.

The invention also provides an isolated polynucleotide encoding thepolypeptide of SEQ ID NO: 126.

Some embodiments of the disclosure also provide an isolated or purifiednucleic acid comprising a polynucleotide which is complementary to thepolynucleotides encoding the CD3ε binding proteins of the disclosure orpolynucleotides which hybridize under stringent conditions to thepolynucleotides encoding the CD3ε binding proteins of the disclosure.

The polynucleotides which hybridize under stringent conditions mayhybridize under high stringency conditions. By “high stringencyconditions” is meant that the polynucleotide specifically hybridizes toa target sequence (the nucleotide sequence of any of the nucleic acidsdescribed herein) in an amount that is detectably stronger thannon-specific hybridization. High stringency conditions includeconditions which would distinguish a polynucleotide with an exactcomplementary sequence, or one containing only a few scatteredmismatches from a random sequence that happened to have a few smallregions (e.g., 3-12 bases) that matched the nucleotide sequence. Suchsmall regions of complementarity are more easily melted than afull-length complement of 14-17 or more bases, and high stringencyhybridization makes them easily distinguishable. Relatively highstringency conditions would include, for example, low salt and/or hightemperature conditions, such as provided by about 0.02-0.1 M NaCl or theequivalent, at temperatures of about 50-70° C. Such high stringencyconditions tolerate little, if any, mismatch between the nucleotidesequence and the template or target strand. It is generally appreciatedthat conditions can be rendered more stringent by the addition ofincreasing amounts of formamide.

The polynucleotide sequences of the disclosure may be operably linked toone or more regulatory elements, such as a promoter or enhancer, thatallow expression of the nucleotide sequence in the intended host cell.The polynucleotide may be a cDNA. The promoter bay be a strong, weak,tissue-specific, inducible or developmental-specific promoter. Exemplarypromoters that may be used are hypoxanthine phosphoribosyl transferase(HPRT), adenosine deaminase, pyruvate kinase, beta-actin, human myosin,human hemoglobin, human muscle creatine, and others. In addition, manyviral promoters function constitutively in eukaryotic cells and aresuitable for use with the described embodiments. Such viral promotersinclude Cytomegalovirus (CMV) immediate early promoter, the early andlate promoters of SV40, the Mouse Mammary Tumor Virus (MMTV) promoter,the long terminal repeats (LTRs) of Maloney leukemia virus, HumanImmunodeficiency Virus (HIV), Epstein Barr Virus (EBV), Rous SarcomaVirus (RSV), and other retroviruses, and the thymidine kinase promoterof Herpes Simplex Virus. Inducible promoters such as the metallothioneinpromoter, tetracycline-inducible promoter, doxycycline-induciblepromoter, promoters that contain one or more interferon-stimulatedresponse elements (ISRE) such as protein kinase R 2′,5′-oligoadenylatesynthetases, Mx genes, ADAR1, and the like may also be sued.

The invention also provides a vector comprising the polynucleotide ofthe invention. The disclosure also provide an expression vectorcomprising the polynucleotide of the invention. Such vectors may beplasmid vectors, viral vectors, vectors for baculovirus expression,transposon based vectors or any other vector suitable for introductionof the synthetic polynucleotide of the invention into a given organismor genetic background by any means. Polynucleotides encoding the CD3εbinding proteins of the disclosure may be operably linked to controlsequences in the expression vector(s) that ensure the expression of theCD3ε binding proteins. Such regulatory elements may include atranscriptional promoter, sequences encoding suitable mRNA ribosomalbinding sites, and sequences that control the termination oftranscription and translation. Expression vectors may also include oneor more nontranscribed elements such as an origin of replication, asuitable promoter and enhancer linked to the gene to be expressed, other5′ or 3′ flanking nontranscribed sequences, 5′ or 3′ nontranslatedsequences (such as necessary ribosome binding sites), a polyadenylationsite, splice donor and acceptor sites, or transcriptional terminationsequences. An origin of replication that confers the ability toreplicate in a host may also be incorporated.

The expression vectors can comprise naturally-occurring ornon-naturally-occurring internucleotide linkages, or both types oflinkages. The non-naturally occurring or altered nucleotides orinternucleotide linkages do not hinder the transcription or replicationof the vector.

Once the vector has been incorporated into the appropriate host, thehost is maintained under conditions suitable for high level expressionof the CD3ε binding proteins of the disclosure encoded by theincorporated polynucleotides. The transcriptional and translationalcontrol sequences in expression vectors to be used in transformingvertebrate cells may be provided by viral sources. Exemplary vectors maybe constructed as described by Okayama and Berg, 3 Mol. Cell. Biol. 280(1983).

Vectors of the disclosure may also contain one or more Internal RibosomeEntry Site(s) (IRES). Inclusion of an IRES sequence into fusion vectorsmay be beneficial for enhancing expression of some proteins. In otherembodiments, the vector system will include one or more polyadenylationsites (e.g., SV40), which may be upstream or downstream of any of theaforementioned nucleic acid sequences. Vector components may becontiguously linked or arranged in a manner that provides optimalspacing for expressing the gene products (i.e., by the introduction of“spacer” nucleotides between the ORFs) or positioned in another way.Regulatory elements, such as the IRES motif, may also be arranged toprovide optimal spacing for expression.

Vectors of the disclosure may be circular or linear. They may beprepared to contain a replication system functional in a prokaryotic oreukaryotic host cell. Replication systems can be derived, e.g., fromColE1, SV40, 2μ plasmid, λ, bovine papilloma virus, and the like.

The recombinant expression vectors can be designed for either transientexpression, for stable expression, or for both. Also, the recombinantexpression vectors can be made for constitutive expression or forinducible expression.

Further, the recombinant expression vectors can be made to include asuicide gene. As used herein, the term “suicide gene” refers to a genethat causes the cell expressing the suicide gene to die. The suicidegene can be a gene that confers sensitivity to an agent, e.g., a drug,upon the cell in which the gene is expressed, and causes the cell to diewhen the cell is contacted with or exposed to the agent. Suicide genesare known in the art and include, for example, the Herpes Simplex Virus(HSV) thymidine kinase (TK) gene, cytosine deaminase, purine nucleosidephosphoryl The vectors may also comprise selection markers, which arewell known in the art. Selection markers include positive and negativeselection marker. Marker genes include biocide resistance, e.g.,resistance to antibiotics, heavy metals, etc., complementation in anauxotrophic host to provide prototrophy, and the like. Exemplary markergenes include antibiotic resistance genes (e.g., neomycin resistancegene, a hygromycin resistance gene, a kanamycin resistance gene, atetracycline resistance gene, a penicillin resistance gene, histidinolresistance gene, histidinol x resistance gene), glutamine synthasegenes, HSV-TK, HSV-TK derivatives for ganciclovir selection, orbacterial purine nucleoside phosphorylase gene for 6-methylpurineselection (Gadi et al., 7 Gene Ther. 1738-1743 (2000)). A nucleic acidsequence encoding a selection marker or the cloning site may be upstreamor downstream of a nucleic acid sequence encoding a polypeptide ofinterest or cloning site.

Exemplary vectors that may be used are Bacterial: pBs, phagescript,PsiX174, pBluescript SK, pBs KS, pNH8a, pNH16a, pNH18a, pNH46a(Stratagene, La Jolla, Calif., USA); pTrc99A, pKK223-3, pKK233-3,pDR540, and pRIT5 (Pharmacia, Uppsala, Sweden). Eukaryotic: pWLneo,pSV2cat, pOG44, PXR1, pSG (Stratagene) pSVK3, pBPV, pMSG and pSVL(Pharmacia), pEE6.4 (Lonza) and pEE12.4 (Lonza). Additional vectorsinclude the pUC series (Fermentas Life Sciences, Glen Burnie, Md.), thepBluescript series (Stratagene, LaJolla, Calif.), the pET series(Novagen, Madison, Wis.), the pGEX series (Pharmacia Biotech, Uppsala,Sweden), and the pEX series (Clontech, Palo Alto, Calif.). Bacteriophagevectors, such as λGT10, λGT11, λEMBL4, and λNM1149, λZapII (Stratagene)can be used. Exemplary plant expression vectors include pBI01, pBI01.2,pBI121, pBI101.3, and pBIN19 (Clontech). Exemplary animal expressionvectors include pEUK-Cl, pMAM, and pMAMneo (Clontech). The expressionvector may be a viral vector, e.g., a retroviral vector, e.g., a gammaretroviral vector.ase, and nitroreductase.

In other embodiments, the vector comprises the polynucleotide encodingthe VH of SEQ ID NO: 55.

In other embodiments, the vector comprises the polynucleotide encodingthe VH of SEQ ID NO: 54.

In other embodiments, the vector comprises the polynucleotide encodingthe VH of SEQ ID NO: 48.

In other embodiments, the vector comprises the polynucleotide encodingthe VL of SEQ ID NO: 59.

In other embodiments, the vector comprises the polynucleotide encodingthe VL of SEQ ID NO: 58.

In other embodiments, the vector comprises the polynucleotide encodingthe VL of SEQ ID NO: 56.

In other embodiments, the vector comprises the polynucleotide encodingthe VH of SEQ ID NO: 55, 54, or 48 and the VL of SEQ ID NOs: 59, 58, or56.

In other embodiments, the vector comprises the polynucleotide encoding

-   -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59;    -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 56;    -   the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 88 and the VL of SEQ ID NO: 58; or    -   the VH of SEQ ID NO: 242 and the VL of SEQ ID NO: 58.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NOs: SEQ ID NOs: 96, 97, 98, 99, 100, 101,102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115,116, 117, 118, 119, 120, 121, 122, 123, 124, 125, or 126.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 96.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 97.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 98.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 99.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 100.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 101.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 102.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 103.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 104.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 105.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 106.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 107.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 108.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 109.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 110.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 112.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 113.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 114.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 115.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 116.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 117.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 118.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 119.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 120.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 121.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 122.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 123.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 124.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 125.

In other embodiments, the vector comprises the polynucleotide encodingthe polypeptide of SEQ ID NO: 126.

The invention also provides for a host cell comprising one or morevectors of the invention. “Host cell” refers to a cell into which avector has been introduced. It is understood that the term host cell isintended to refer not only to the particular subject cell but to theprogeny of such a cell, and also to a stable cell line generated fromthe particular subject cell. Because certain modifications may occur insucceeding generations due to either mutation or environmentalinfluences, such progeny may not be identical to the parent cell, butare still included within the scope of the term “host cell” as usedherein. Such host cells may be eukaryotic cells, prokaryotic cells,plant cells or archeal cells. Escherichia coli, bacilli, such asBacillus subtilis, and other enterobacteriaceae, such as Salmonella,Serratia, and various Pseudomonas species are examples of prokaryotichost cells. Other microbes, such as yeast, are also useful forexpression. Saccharomyces (e.g., S. cerevisiae) and Pichia are examplesof suitable yeast host cells. Exemplary eukaryotic cells may be ofmammalian, insect, avian or other animal origins. Mammalian eukaryoticcells include immortalized cell lines such as hybridomas or myeloma celllines such as SP2/0 (American Type Culture Collection (ATCC), Manassas,Va., CRL-1581), NS0 (European Collection of Cell Cultures (ECACC),Salisbury, Wiltshire, UK, ECACC No. 85110503), FO (ATCC CRL-1646) andAg653 (ATCC CRL-1580) murine cell lines. An exemplary human myeloma cellline is U266 (ATTC CRL-TIB-196). Other useful cell lines include thosederived from Chinese Hamster Ovary (CHO) cells such as CHO-K1SV (LonzaBiologics, Walkersville, Md.), CHO-K1 (ATCC CRL-61) or DG44.

The disclosure also provides a method of producing the CD3ε bindingprotein of the disclosure comprising culturing the host cell of thedisclosure in conditions that the CD3ε binding protein is expressed, andrecovering the CD3ε binding protein produced by the host cell. Methodsof making proteins and purifying them are known. Once synthesized(either chemically or recombinantly), the CD3ε binding proteins may bepurified according to standard procedures, including ammonium sulfateprecipitation, affinity columns, column chromatography, high performanceliquid chromatography (HPLC) purification, gel electrophoresis, and thelike (see generally Scopes, Protein Purification (Springer-Verlag, N.Y.,(1982)). A subject protein may be substantially pure, e.g., at leastabout 80% to 85% pure, at least about 85% to 90% pure, at least about90% to 95% pure, or at least about 98% to 99%, or more, pure, e.g., freefrom contaminants such as cell debris, macromolecules, etc. other thanthe subject protein

The polynucleotides encoding the CD3ε binding proteins of the disclosuremay be incorporated into vectors using standard molecular biologymethods. Host cell transformation, culture, antibody expression andpurification are done using well known methods.

Modified nucleotides may be used to generate the polynucleotides of thedisclosure. Exemplary modified nucleotides are 5-fluorouracil,5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine,4-acetylcytosine, 5-(carboxyhydroxymethyl) uracil,carboxymethylaminomethyl-2-thiouridine,5-carboxymethylaminomethyluracil, dihydrouracil, N⁶-substituted adenine,7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine,5″-methoxycarboxymethyluracil, 5-methoxyuracil,2-methylthio-N⁶-isopentenyladenine, uracil-5-oxyacetic acid (v),wybutoxosine, pseudouracil, queuosine, beta-D-galactosylqueosine,inosine, N⁶-isopentenyladenine, 1-methylguanine, 1-methylinosine,2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine,5-methylcytosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil,4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester,3-(3-amino-3-N-2-carboxypropyl) uracil, and 2,6-diaminopurine.

Pharmaceutical Compositions/Administration

The disclosure also provides a pharmaceutical composition comprising theCD3ε binding protein of the disclosure and a pharmaceutically acceptablecarrier.

The disclosure also provides a pharmaceutical composition comprising theantigen binding domain that binds CD3ε of the disclosure and apharmaceutically acceptable carrier.

The disclosure also provides a pharmaceutical composition comprising theprotein comprising the antigen binding domain that binds CD3ε of thedisclosure and a pharmaceutically acceptable carrier.

The disclosure also provides a pharmaceutical composition comprising themultispecific protein comprising the antigen binding domain that bindsCD3ε of the disclosure and a pharmaceutically acceptable carrier.

The disclosure also provides a pharmaceutical composition comprising themultispecific protein comprising the antigen binding domain that bindsCD3ε and antigen binding domain that binds a tumor antigen of thedisclosure and a pharmaceutically acceptable carrier.

For therapeutic use, the CD3ε binding protein of the disclosure may beprepared as pharmaceutical compositions containing an effective amountof the antibody as an active ingredient in a pharmaceutically acceptablecarrier. These solutions are sterile and generally free of particulatematter. They may be sterilized by conventional, well-known sterilizationtechniques (e.g., filtration). The compositions may containpharmaceutically acceptable auxiliary substances as required toapproximate physiological conditions such as pH adjusting and bufferingagents, stabilizing, thickening, lubricating and coloring agents, etc.

The term “pharmaceutically acceptable,” as used herein with regard topharmaceutical compositions, means approved by a regulatory agency ofthe Federal or a state government or listed in the U.S. Pharmacopeia orother generally recognized pharmacopeia for use in animals and/or inhumans.

Methods of Treatment and Uses

The disclosure also provides the bispecific or multispecific proteincomprising a first antigen biding domain that specifically binds CD3εand a second antigen biding domain that specifically binds a secondantigen of the disclosure for use in therapy.

The disclosure also provides the bispecific or multispecific proteincomprising a first antigen biding domain that specifically binds CD3εand a second antigen biding domain that specifically binds a secondantigen of the disclosure for use in treating a cell proliferativedisorder.

The disclosure also provides the bispecific or multispecific proteincomprising a first antigen biding domain that specifically binds CD3εand a second antigen biding domain that specifically binds a secondantigen of the disclosure for use in killing cancer cells.

The disclosure also provides the bispecific or multispecific proteincomprising a first antigen biding domain that specifically binds CD3εand a second antigen biding domain that specifically binds a secondantigen of the disclosure for use in the manufacture of a medicament forkilling cancer cells.

The disclosure also provides the bispecific or multispecific proteincomprising a first antigen biding domain that specifically binds CD3εand a second antigen biding domain that specifically binds a secondantigen of the disclosure for use in redirection of cytolytic T cells.

The disclosure also provides the bispecific or multispecific proteincomprising a first antigen biding domain that specifically binds CD3εand a second antigen biding domain that specifically binds a secondantigen of the disclosure for use in the manufacture of a medicament forredirection of cytolytic T cells.

The disclosure also provides the bispecific or multispecific proteincomprising a first antigen biding domain that specifically binds CD3εand a second antigen biding domain that specifically binds a secondantigen of the disclosure for use in redirection of cytolytic T cells inthe tumor microenviroment.

The disclosure also provides the bispecific or multispecific proteincomprising a first antigen biding domain that specifically binds CD3εand a second antigen biding domain that specifically binds a secondantigen of the disclosure for use in the manufacture of a medicament forredirection of cytolytic T cells in the tumor microenviroment.

The disclosure also provides the bispecific or multispecific proteincomprising a first antigen biding domain that specifically binds CD3εand a second antigen biding domain that specifically binds a secondantigen of the disclosure for use in treating cancer.

The disclosure also provides the bispecific or multispecific proteincomprising a first antigen biding domain that specifically binds CD3εand a second antigen biding domain that specifically binds a secondantigen of the disclosure for use in the manufacture of a medicament fortreating cancer.

In one aspect, the disclosure relates generally to the treatment of asubject at risk of developing cancer. The invention also includestreating a malignancy in which chemotherapy and/or immunotherapy resultsin significant immunosuppression in a subject, thereby increasing therisk of the subject developing cancer.

The disclosure also provides a method of treating a noncancerouscondition in a subject at risk of developing a cancerous condition,comprising administering the antigen binding domain that bind CD3ε ofthe disclosure to the subject to treat the noncancerous condition.

The disclosure also provides a method of treating a noncancerouscondition in a subject at risk of developing a cancerous condition,comprising administering the protein comprising the antigen bindingdomain that bind CD3ε of the disclosure to the subject to treat thenoncancerous condition.

The disclosure also provides a method of treating a noncancerouscondition in a subject at risk of developing a cancerous condition,comprising administering the multispecific protein comprising theantigen binding domain that bind CD3ε of the disclosure to the subjectto treat the noncancerous condition.

The disclosure also provides a method of treating a noncancerouscondition in a subject at risk of developing a cancerous condition,comprising administering the immunoconjugate of the disclosure to thesubject to treat the noncancerous condition.

The disclosure also provides a method of treating a noncancerouscondition in a subject at risk of developing a cancerous condition,comprising administering the pharmaceutical composition of thedisclosure to the subject to treat the noncancerous condition.

The disclosure also provides a method of treating cancer in a subject,comprising administering a therapeutically effective amount of themultispecific protein comprising the antigen binding domain that bindsCD3ε to the subject to treat the cancer, wherein the antigen bindingdomain that bind CD3ε comprises

-   -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59;    -   the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 56;    -   the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 58;    -   the VH of SEQ ID NO: 88 and the VL of SEQ ID NO: 58; or    -   the VH of SEQ ID NO: 242 and the VL of SEQ ID NO: 58.

The disclosure also provides a method of treating cancer in a subject,comprising administering a therapeutically effective amount of themultispecific protein comprising the antigen binding domain that bindsCD3ε to the subject to treat the cancer, wherein the antigen bindingdomain that binds CD3ε comprises the amino acid sequence of SEQ ID NOs:96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110,111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124,125, or 126.

A further aspect of the disclosure is a method of treating a cellproliferative disorder in a subject in need thereof, the methodcomprising administering to the subject a therapeutically effectiveamount of the bispecific or multispecific protein comprising a firstantigen biding domain that specifically binds CD3ε and a second antigenbiding domain that specifically binds a second antigen of thedisclosure. In other embodiments, the bispecific or multispecificprotein comprising a first antigen biding domain that specifically bindsCD3ε and a second antigen biding domain that specifically binds a secondantigen of the disclosure, is administered to the subject.

In any of the preceding uses or methods, the cell proliferative disorderis cancer. In other embodiments, the cancer is selected from the groupconsisting of esophageal cancer, stomach cancer, small intestine cancer,large intestine cancer, colorectal cancer, breast cancer, non-small celllung cancer, non-Hodgkin's lymphoma (NHL), B cell lymphoma, B cellleukemia, multiple myeloma, renal cancer, prostate cancer, liver cancer,head and neck cancer, melanoma, ovarian cancer, mesothelioma,glioblastoma, germinal-center B-cell-like (GCB) DLBCL, activatedB-cell-like (ABC) DLBCL, follicular lymphoma (FL), mantle cell lymphoma(MCL), acute myeloid leukemia (AML), chronic lymphoid leukemia (CLL),marginal zone lymphoma (MZL), small lymphocytic leukemia (SLL),lymphoplasmacytic lymphoma (LL), Waldenstrom macroglobulinemia (WM),central nervous system lymphoma (CNSL), Burkitt's lymphoma (BL), B-cellprolymphocytic leukemia, Splenic marginal zone lymphoma, Hairy cellleukemia, Splenic lymphoma/leukemia, unclassifiable, Splenic diffuse redpulp small B-cell lymphoma, Hairy cell leukemia variant, Waldenstrommacroglobulinemia, Heavy chain diseases, Plasma cell myeloma, Solitaryplasmacytoma of bone, Extraosseous plasmacytoma, Extranodal marginalzone lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma),Nodal marginal zone lymphoma, Pediatric nodal marginal zone lymphoma,Pediatric follicular lymphoma, Primary cutaneous follicle centrelymphoma, T-cell/histiocyte rich large B-cell lymphoma, Primary DLBCL ofthe CNS, Primary cutaneous DLBCL, leg type, EBV-positive DLBCL of theelderly, DLBCL associated with chronic inflammation, Lymphomatoidgranulomatosis, Primary mediastinal (thymic) large B-cell lymphoma.Intravascular large B-cell lymphoma, ALK-positive large B-cell lymphoma,Plasmablastic lymphoma, Large B-cell lymphoma arising in HHV8-associatedmulticentric Castleman disease, Primary effusion lymphoma: B-celllymphoma, unclassifiable, with features intermediate between diffuselarge B-cell lymphoma and Burkitt lymphoma, and B-cell lymphoma,unclassifiable, with features intermediate between diffuse large B-celllymphoma, classical Hodgkin lymphoma and light chain amyloidosis.

In other embodiments, the cancer is esophageal cancer. In otherembodiments, the cancer is an adenocarcinoma, for example, a metastaticadenocarcinoma (e.g., a colorectal adenocarcinoma, a gastricadenocarcinoma, or a pancreatic adenocarcinoma).

In another aspect, the disclosure features a kit comprising: (a) acomposition comprising any one of the preceding the bispecific ormultispecific protein comprising a first antigen biding domain thatspecifically binds CD3ε and a second antigen biding domain thatspecifically binds a second antigen of the disclosure and (b) a packageinsert comprising instructions for administering the composition to asubject to treat or delay progression of a cell proliferative disorder.

In any of the preceding uses or methods, the subject can be a human.

Combination Therapies

The CD3ε binding proteins of the disclosure may be administered incombination with at least one additional therapeutics.

In other embodiments, the delivery of one treatment is still occurringwhen the delivery of the second begins, so that there is overlap interms of administration. This is sometimes referred to herein as“simultaneous” or “concurrent delivery”. In other embodiments, thedelivery of one treatment ends before the delivery of the othertreatment begins. In some embodiments of either case, the treatment ismore effective because of combined administration. For example, thesecond treatment is more effective, e.g., an equivalent effect is seenwith less of the second treatment, or the second treatment reducessymptoms to a greater extent, than would be seen if the second treatmentwere administered in the absence of the first treatment, or theanalogous situation is seen with the first treatment. In otherembodiments, delivery is such that the reduction in a symptom, or otherparameter related to the disorder is greater than what would be observedwith one treatment delivered in the absence of the other. The deliverycan be such that an effect of the first treatment delivered is stilldetectable when the second is delivered.

The CD3ε binding proteins described herein and the at least oneadditional therapeutic agent can be administered simultaneously, in thesame or in separate compositions, or sequentially. For sequentialadministration, the CD3ε binding proteins described herein can beadministered first, and the additional agent can be administered second,or the order of administration can be reversed.

Embodiments

This invention provides the following non-limiting embodiments.

-   -   1. An isolated protein comprising an antigen binding domain that        binds to cluster of differentiation 3ε (CD3ε), wherein the        antigen binding domain that binds CD3ε comprises:        -   a. a heavy chain complementarity determining region (HCDR)            1, a HCDR2 and a HCDR3 of a heavy chain variable region (VH)            of SEQ ID NO: 55 and a light chain complementarity            determining region (LCDR) 1, a LCDR2 and a LCDR3 of a light            chain variable region (VL) of SEQ ID NO: 59;        -   b. the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID            NO: 55 and the LCDR1, the LCDR2 and the LCDR3 of the VL of            SEQ ID NO: 58;        -   c. the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID            NO: 54 and the LCDR1, the LCDR2 and the LCDR3 of the VL of            SEQ ID NO: 56; or        -   d. the HCDR1, the HCDR2 and the HCDR3 of the VH of SEQ ID            NO: 48 and the LCDR1, the LCDR2 and the LCDR3 of the VL of            SEQ ID NO: 58;    -    wherein the amino acid in position N106 of SEQ ID NO: 55, 54,        or 48 is optionally substituted with the amino acid selected        from the group consisting of A, G, S, F, E, T, R, V, I, Y, L, P,        Q, and K, wherein the residue numbering starts from N-terminus        of SEQ ID NO: 55, 54, or 48.    -   2. An isolated protein, comprising the HCDR1, the HCDR2, the        HCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 70, 71,        86, 79, 80, and 81, respectively.    -   3. The isolated protein of embodiment 1 or 2, comprising the        HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and the LCDR3        of        -   a. SEQ ID NOs: 70, 71, 72, 79, 80, and 81, respectively;        -   b. SEQ ID NOs: 70, 71, 87, 79, 80, and 81, respectively; or        -   c. SEQ ID NOs: 70, 71, 90, 79, 80, and 81, respectively.    -   4. The isolated protein of embodiments 1-3, wherein the antigen        binding domain that binds CD3ε is a scFv, a (scFv)2, a Fv, a        Fab, a F(ab′)2, a Fd, a dAb or a VHH.    -   5. The isolated protein of embodiment 4, wherein the antigen        binding domain that binds CD3ε is the Fab.    -   6. The isolated protein of embodiment 4, wherein the antigen        binding domain that binds CD3ε is the scFv.    -   7. The isolated protein of embodiment 6, wherein the scFv        comprises, from the N- to C-terminus, a VH, a first linker (L1)        and a VL (VH-L1-VL) or the VL, the L1 and the VH (VL-L1-VH).    -   8. The isolated protein of embodiment 7, wherein the L1        comprises        -   a. about 5-50 amino acids;        -   b. about 5-40 amino acids;        -   c. about 10-30 amino acids; or        -   d. about 10-20 amino acids.    -   9. The isolated protein of embodiment 7, wherein the L1        comprises an amino acid sequence of SEQ ID NOs: 3-36.    -   10. The isolated protein of embodiment 9 wherein the L1        comprises the amino acid sequence of SEQ ID NO: 3.    -   11. The isolated protein of any one of embodiments 1-10, wherein        the antigen binding domain that binds CD3ε comprises the VH of        SEQ ID NOs: 55, 54, or 48 and the VL of SEQ ID NOs: 59, 58 or        56.    -   12. The isolated protein of embodiment 11, wherein the antigen        binding domain that binds CD3ε comprises:        -   a. the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59;        -   b. the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58;        -   c. the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 56;        -   d. the VH of SEQ ID NO: 48 and the VL of SEQ ID NO: 58;        -   e. the VH of SEQ ID NO: 88 and the VL of SEQ ID NO: 58; or        -   f. the VH of SEQ ID NO: 242 and the VL of SEQ ID NO: 58.    -   13. The isolated protein of any one of embodiments 1-12, wherein        the antigen binding domain that binds CD3ε comprises an amino        acid sequence selected from the group consisting of SEQ ID NOs:        96-126.    -   14. The isolated protein of any one of embodiments 1-13, wherein        the isolated protein is a multispecific protein.    -   15. The isolated protein of embodiment 14, wherein the        multispecific protein is a bispecific protein.    -   16. The isolated protein of embodiment 14, wherein the        multispecific protein is a trispecific protein.    -   17. The isolated protein of any one of embodiments 1-16, further        comprising an immunoglobulin (Ig) constant region or a fragment        of the Ig constant region thereof    -   18. The isolated protein of embodiment 17, wherein the fragment        of the Ig constant region comprises a Fc region.    -   19. The isolated protein of embodiment 17, wherein the fragment        of the Ig constant region comprises a CH2 domain.    -   20. The isolated protein of embodiment 17, wherein the fragment        of the Ig constant region comprises a CH3 domain.    -   21. The isolated protein of embodiment 17, wherein the fragment        of the Ig constant region comprises a CH2 domain and a CH3        domain.    -   22. The isolated protein of embodiment 17, wherein the fragment        of the Ig constant region comprises at least portion of a hinge,        a CH2 domain and a CH3 domain.    -   23. The isolated protein of embodiment 17, wherein the fragment        of the Ig constant region comprises a hinge, a CH2 domain and a        CH3 domain.    -   24. The isolated protein of any one of embodiments 17-24,        wherein the antigen binding domain that binds CD3ε is conjugated        to the N-terminus of the Ig constant region or the fragment of        the Ig constant region.    -   25. The isolated protein of any one of embodiments 17-24,        wherein the antigen binding domain that binds CD3ε is conjugated        to the C-terminus of the Ig constant region or the fragment of        the Ig constant region.    -   26. The isolated protein of any one of embodiments 17-24,        wherein the antigen binding domain that binds CD3ε is conjugated        to the Ig constant region or the fragment of the Ig constant        region via a second linker (L2).    -   27. The isolated protein of embodiment 35, wherein the L2        comprises the amino acid sequence selected from the group        consisting of SEQ ID NOs: 3-36.    -   28. The isolated protein of any one of embodiments 14-27,        wherein the multispecific protein comprises an antigen binding        domain that binds an antigen other than CD3ε.    -   29. The multispecific antibody of embodiment 14-28, wherein the        cell antigen is a tumor associated antigen.    -   30. The isolated protein of any one of embodiments 14-29,        wherein the Ig constant region or the fragment of the Ig        constant region is an IgG1, an IgG2, an IgG3 or an IgG4 isotype.    -   31. The isolated protein of any one of embodiments 1-30, wherein        the Ig constant region or the fragment of the Ig constant region        comprises at least one mutation that results in reduced binding        of the protein to a Fcγ receptor (FcγR).    -   32. The isolated protein of embodiment 31, wherein the at least        one mutation that results in reduced binding of the protein to        the FcγR is selected from the group consisting of F234A/L235A,        L234A/L235A, L234A/L235A/D265S,        V234A/G237A/P238S/H268A/V309L/A330S/P331S, F234A/L235A,        S228P/F234A/L235A, N297A, V234A/G237A,        K214T/E233P/L234V/L235A/G236-deleted/A327G/P331A/D365E/L358M,        H268Q/V309L/A330S/P331S, S267E/L328F, L234F/L235E/D265A,        L234A/L235A/G237A/P238S/H268A/A330S/P331S,        S228P/F234A/L235A/G237A/P238S and        S228P/F234A/L235A/G236-deleted/G237A/P238S, wherein residue        numbering is according to the EU index.    -   33. The isolated protein of any one of embodiments 31-32,        wherein the FcγR is FcγRI, FcγRIIA, FcγRIIB or FcγRIII, or any        combination thereof.    -   34. The isolated protein of any one of the embodiments 14-33,        wherein the protein comprises at least one mutation in a CH3        domain of the Ig constant region.    -   35. The isolated protein of embodiment 34, wherein the at least        one mutation in the CH3 domain of the Ig constant region is        selected from the group consisting of T350V, L351Y, F405A,        Y407V, T366Y, T366W, T366L, T366L, F405W, T394W, K392L, T394S,        T394W, Y407T, Y407A, T366S/L368A/Y407V, L351Y/F405A/Y407V,        T366I/K392M/T394W, F405A/Y407V, T366L/K392M/T394W,        T366L/K392L/T394W, L351Y/Y407A, L351Y/Y407V, T366A/K409F,        T366V/K409F, T366A/K409F, T350V/L351Y/F405A/Y407V and        T350V/T366L/K392L/T394W, wherein residue numbering is according        to the EU index.    -   36. A pharmaceutical composition comprising the isolated protein        of any one of embodiments 1-35 and a pharmaceutically acceptable        carrier.    -   37. A polynucleotide encoding the isolated protein of any one of        embodiments 1-35.    -   38. A vector comprising the polynucleotide of embodiment 35.    -   39. A host cell comprising the vector of embodiment 38.    -   40. A method of producing the isolated protein of any one of        embodiments 1-35, comprising culturing the host cell of        embodiment 39 in conditions that the protein is expressed, and        recovering the protein produced by the host cell.    -   41. A method of treating a cancer in a subject, comprising        administering a therapeutically effective amount of the isolated        protein of any one of embodiments 1-35 to the subject in need        thereof to treat the cancer.    -   42. An anti-idiotypic antibody binding to the isolated protein        of any one of embodiments 1-35.    -   43. An isolated protein of any one of embodiments 1-35        comprising an amino acid sequence selected from the group        consisting of SEQ ID NOs: 127-157.    -   44. An isolated protein of any one of embodiments 1-35        comprising an antibody heavy chain of SEQ ID NO: 224 and        antibody light chain of SEQ ID NO: 226.

EXAMPLES Example 1. Generation and Characterization of Anti-CD3 mAbs

The publicly available mouse Cris7 antibody, specific to human CD3ε(Alberola-Ila, J. et al. Stimulation through the TCR/CD3 complexup-regulates the CD2 surface expression on human T lymphocytes. JImmunol 146, 1085-1092 (1991)) was used for these experimentations. TheVH and VL sequences of Cris-7 are shown below.

Cris-7 VH (SEQ ID NO: 37):

QVQLQQSGAELARPGASVKMSCKASGYTFTRSTMHWVKQRPGQGLEWIGYINPSSAYTNYNQKFKDKATLTADKSSSTAYMQLSSLTSEDSAVYYCASPQVHYDYNGFPYWGQGTLVTVSA

Cris-7 VL (SEQ ID NO: 38):

QVVLTQSPAIMSAFPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDSSKLASGVPARFSGSGSGTSYSLTISSMETEDAATYYCQQWSRNPPTFGGGTKLQIT

Humanization and scFv Formatting of CD3 Binding Domains

Evaluation of Optimal Germline Sequences

Murine Cris-7 was humanized in the single-chain fragment variable-domain(scFv) format. To find the binding affinity matched to Cris7 and themost thermal-stable combination of human germline acceptor HC and LCpair for scFv format, two human heavy variable-domain (Hv) germlinesequences and two human light variable-domain (Lv) germline sequenceswere selected for the antibody humanization: IGHV1-69*02-IGHJ1-01 andIGHV5-10-1*01-IIGHJ1-01 for Hv and IGKV3-11*02-IGKJ4-01 or theIGKV1-39*01-IGKJ4-01 germline for Lv (Retrieved from the Internet: <URL:http://www.imgt.org/vquest/refseqh.html>). The CDR-grafted sequenceswere generated with limited back mutations to enhance stability (seeTable 4 below). These CDR-grafted v-regions were then expressed in E.coli in scFv format in both the heavy chain-linker-light chain (HL) andin the light chain-linker-heavy chain (LH) orientations. A matrix of Hvand Lv pairings was evaluated in scFv-format in both orientations of Lvfollowed by Hv or Hv followed by Lv with a flexible linker between thesevariable domains, as described below. CD3B1127 and CD3B1128 comprisedmurine VH and VL sequences (Table 4). There were two main conclusionsfrom this experiment. First, in all cases, the Cris-7-derived scFvmolecules displayed significantly stronger binding to recombinant CD3(TRCW5, SEQ ID NO: 39) in the HL orientation compared to the LHorientation, based primarily on higher maximum signal, as determined byELISA. Second, the IGHV1-69*02-IGHJ1-01 heavy chain germline withIGKV3-11*02-IGKJ4-01 light chain germline grafted construct containinglimited back mutations in the heavy-light orientation exhibited the bestexpression, binding profile, and potential differentiation, and so waschosen for humanization (FIG. 1 , Tables 4 and 5).

TABLE 4Amino acid sequences of grafted sequences, comprising limited back mutations.Protein ID VH VL Linker (orientation) (SEQ ID NO:) (SEQ ID NO: )(SEQ ID NO:) Cris-7 QVQLQQSGAELARPGASVKMS QVVLTQSPAIMSAFPGEKV GGSEGKSSGCKASGYTFTRSTMHWVKQRPG TMTCSASSSVSYMNWYQQ SGSESKSTGQGLEWIGYINPS SAYTNYNQKF KSGTSPKRWIYDSSKLASG GS KDKATLTADKSSSTAYMQLSSLVPARFSGSGSGTSYSLTISS (3) TSEDSAVYYCASPQVHYDYNG METEDAATYYCQQWSRNPFPYWGQGTLVTVSA PTFGGGTKLQIT (37) (38) CD3B1129 QVQLVQSGAEVKKPGSSVKVSDIQLTQSPSSLSASVGDRV GGSEGKSSG (HL*) CKASGYTFTRSTMHWVRQAPGTITCSASSSVSYMNWYQQ SGSESKSTG QGLEWMGYINPSSAYTNYNQK KPGTSPKRLIYDSSKLASGGS FQGRVTLTADKSTSTAYMELSS VPSRFSGSGSGTDYTLTISS (3) LRSEDTAVYYCARPQVHYDYNLQPEDFATYYCQQWSRNP GFPYWGQGTLVTVSS PTFGGGTKVEIK (40) (42) CD3B1130QVQLVQSGAEVKKPGSSVKVS EIVLTQSPATLSLSPGERAT GGSEGKSSG (HL)CKASGYTFTRSTMHWVRQAPG LSCSASSSVSYMNWYQQK SGSESKSTG QGLEWMGYINPSSAYTNYNQKPGTSPRRLIYDSSKLASGIP GS FQGRVTLTADKSTSTAYMELSS ARFSGSGSGRDYTLTISSLE (3)LRSEDTAVYYCARPQVHYDYN PEDFAVYYCQQWSRNPPT GFPYWGQGTLVTVSS FGGGTKVEIK (40)(43) CD3B1131 EVQLVQSGAEVKKPGESLRISC DIQLTQSPSSLSASVGDRV GGSEGKSSG (HL)KASGYTFTRSTMHWVRQMPG TITCSASSSVSYMNWYQQ SGSESKSTG KGLEWMGYINPSSAYTNYNPSFKPGTSPKRLIYDSSKLASG GS QGHVTLSADKSISTAYLQWSSL VPSRFSGSGSGTDYTLTISS (3)KASDTAMYYCARPQVHYDYN LQPEDFATYYCQQWSRNP GFPYWGQGTLVTVSS PTFGGGTKVEIK(41) (42) CD3B1132 EVQLVQSGAEVKKPGESLRISC EIVLTQSPATLSLSPGERAT GGSEGKSSG(HL) KASGYTFTRSTMHWVRQMPG LSCSASSSVSYMNWYQQK SGSESKSTGKGLEWMGYINPSSAYTNYNPSF PGTSPRRLIYDSSKLASGIP GS QGHVTLSADKSISTAYLQWSSLARFSGSGSGRDYTLTISSLE (3) KASDTAMYYCARPQVHYDYN PEDFAVYYCQQWSRNPPTGFPYWGQGTLVTVSS FGGGTKVEIK (41) (43) CD3B1133 QVQLVQSGAEVKKPGSSVKVSDIQLTQSPSSLSASVGDRV GGSEGKSSG (LH) CKASGYTFTRSTMHWVRQAPGTITCSASSSVSYMNWYQQ SGSESKSTG QGLEWMGYINPSSAYTNYNQK KPGTSPKRLIYDSSKLASGGS FQGRVTLTADKSTSTAYMELSS VPSRFSGSGSGTDYTLTISS (3) LRSEDTAVYYCARPQVHYDYNLQPEDFATYYCQQWSRNP GFPYWGQGTLVTVSS PTFGGGTKVEIK (40) (42) CD3B1134EVQLVQSGAEVKKPGESLRISC DIQLTQSPSSLSASVGDRV GGSEGKSSG (LH)KASGYTFTRSTMHWVRQMPG TITCSASSSVSYMNWYQQ SGSESKSTG KGLEWMGYINPSSAYTNYNPSFKPGTSPKRLIYDSSKLASG GS QGHVTLSADKSISTAYLQWSSL VPSRFSGSGSGTDYTLTISS (3)KASDTAMYYCARPQVHYDYN LQPEDFATYYCQQWSRNP GFPYWGQGTLVTVSS PTFGGGTKVEIK(41) (42) CD3B1135 QVQLVQSGAEVKKPGSSVKVS EIVLTQSPATLSLSPGERAT GGSEGKSSG(LH) CKASGYTFTRSTMHWVRQAPG LSCSASSSVSYMNWYQQK SGSESKSTGQGLEWMGYINPSSAYTNYNQK PGTSPRRLIYDSSKLASGIP GS FQGRVTLTADKSTSTAYMELSSARFSGSGSGRDYTLTISSLE (3) LRSEDTAVYYCARPQVHYDYN PEDFAVYYCQQWSRNPPTGFPYWGQGTLVTVSS FGGGTKVEIK (40) (43) CD3B1136 EVQLVQSGAEVKKPGESLRISCEIVLTQSPATLSLSPGERAT GGSEGKSSG (LH) KASGYTFTRSTMHWVRQMPGLSCSASSSVSYMNWYQQK SGSESKSTG KGLEWMGYINPSSAYTNYNPSF PGTSPRRLIYDSSKLASGIPGS QGHVTLSADKSISTAYLQWSSL ARFSGSGSGRDYTLTISSLE (3) KASDTAMYYCARPQVHYDYNPEDFAVYYCQQWSRNPPT GFPYWGQGTLVTVS S FGGGTKVEIK (41) (43) CD3B1127QVQLQQSGAELARPGASVKMS QVVLTQSPAIMSAFPGEKV GGSEGKSSG (HL)CKASGYTFTRSTMHWVKQRPG TMTCSASSSVSYMNWYQQ SGSESKSTGQGLEWIGYINPS SAYTNYNQKF KSGTSPKRWIYDSSKLASG GS KDKATLTADKSSSTAYMQLSSLVPARFSGSGSGTSYSLTISS (3) TSEDSAVYYCASPQVHYDYNG METEDAATYYCQQWSRNPFPYWGQGTLVTVSA PTFGGGTKLQIT (37) (38) CD3B1128 QVQLQQSGAELARPGASVKMSQVVLTQSPAIMSAFPGEKV GGSEGKSSG (LH) CKASGYTFTRSTMHWVKQRPGTMTCSASSSVSYMNWYQQ SGSESKSTG QGLEWIGYINPS SAYTNYNQKF KSGTSPKRWIYDSSKLASGGS KDKATLTADKSSSTAYMQLSSL VPARFSGSGSGTSYSLTISS (3) TSEDSAVYYCASPQVHYDYNGMETEDAATYYCQQWSRNP FPYWGQGTLVTVSA PTFGGGTKLQIT (37) (38) *HL-VH-Linker-VL; LH-VL-Linker-VH

TABLE 5 EC50 (nM) for binding of the CD3-specific variants torecombinant CD3, using ELISA. Protein EC50 CD3B1129 0.7124 CD3B11300.7465 CD381131 1.137 CD3B1132 ~1.101 CD3B1133 0.9583 CD3B1134 ~0.006296CD3B1135 1.036 CD3B1136 CD3B1127 ~0.3972 CD3B1128 ~0.4369 F5 HL~0.005701 Media

Human Framework Optimization in the IGHV1-69*02-IGHJ1-01 andIGKV3-11*02-IGKJ4-01 Germline

Since the IGHV1-69*02-IGHJ1-01 and IGKV3-11*02-IGKJ4-01 germline graftedsequences (CD3B1130) displayed enhanced binding compared to the murineparents and represented the human germline most similar to the murineparent, as described above, human framework adaptation was performedstarting from this CDR-grafted sequence. Starting from this sequence,several sites in the VH were selected which may influence stability ofthe molecule were identified and were thus selected for library-basedmouse back-mutagenesis (Table 6). In one VH library, 4 sites (M48I,A60N, V67A, and I69L—Kabat numbering) were mutated in binary librariesand R94 (Kabat numbering) was mutated to S, V, L, K, T, R, I, or Y for atotal of 128 variants. In a second library, 9 sites (K12A, V20M, R38K,M48I, A60N, R66, V67A, I69L, and R94S—Kabat numbering) were mutated in abinary library for a total of 512 variants. These methods are known inthe art and is described, for example, in Chiu et al., Antibodies 2019,8, 55.

TABLE 6Murine Cris-7, human Germline VH sequences used for humanization,and position of binary and mutated residues. Name VH Cris-7 VHQVQLQQSGAELARPGASVKMSCKASGYTFTRSTMHWVKQRPGQG LEW I GYINPSSAYTNY NQKFKDKAT L T A DKSSSTAYMQLSSLTSED SAVYYCA

PQVHYDYNGFPYWGQGTLVTVSA (SEQ ID NO: 37) IGHV1-69*02-IGHJ1-QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYTISWVRQAPGQGLE 01 W M GRIIPILGIANY AQKFQGR V T I TADKSTSTAYMELSSLRSEDTAV Library 1 YYCA

xxxWGQGTTVTVSS (SEQ ID NO: 44) IGHV1-69*02-IGHJ1- QVQLVQSGAEV K KPGSSVKV SCKASGGTFSSYTISWV R QAPGQGLE 01 W M GRIIPILGIANY A QKFQG RV T ITADKSTSTAYMELSSLRSEDTAV Library 2 YYCA

xxxWGQGTTVTVSS (SEQ ID NO: 45)

Analogously, two libraries were generated for the VL sequence, byidentifying sites which may influence stability of the molecule. Inlibrary 1, no changes were made to the LC. In library 2, 11 sites wereselected for mouse back-mutagenesis in binary fashion (L11M, L13A, A19V,L21M, Q42T, A43S, L46R, L47W, I58V, F71Y, and L78M) for a total of 2048variants (Table 7).

TABLE 7Murine Cris-7, human Germline VL sequences used for humanization,and position of binary and mutated residues. Name VL Cris-7 VLQVVLTQSPAI M S A FPGEK V T M TCSASSSVSYMNWYQQKSG TS PK R W IYDSSKLASG VPARFSGSGSGTS Y SLTISS M ETEDAATYYCQQWS RNPPTFGGGTKLQIT (SEQ ID NO: 38)IGKV3-11*02-IGKJ4- EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLL 01IYDASNRATGIPARFSGSGSGRDFTLTISSLEPEDFAVYYCQQRSNWP Library 1 XXXFGGGTKVEIK(SEQ ID NO: 46) IGKV3-11*02-IGKJ4- EIVLTQSPAT L S L SPGER A T LSCRASQSVSSYLAWYQQKPG Q A PR L 01 L IYDASNRATG I PARFSGSGSGRD F TLTISS LEPEDFAVYYCQQRSN Library 2 WPXXXFGGGTKVEIK (SEQ ID NO: 47)

Back mutation libraries were created through molecular biologytechniques known in the art (Thomas S., et al. DNA library constructionusing Gibson Assembly®. Nat Methods, p.i-ii November 2015). Differentacceptor germlines were paired with the opposite murine parent chain. Inthis manner, only one chain was human framework adapted with potentialback mutations at a single time.

Briefly, DNA was transformed into an E. coli expression vector togenerate scFv molecules having a C-terminal HA-tag, and cells wereplated on 2×YT/Carb/2% Glucose grown overnight at 37 C. Colonies werepicked and transferred 50ul of overnight growth cultures to new platescontaining 500ul of 2×YT/Carb/0.1% Glucose, grown for 6-7 hr, andcombined with 50ul 2×YT medium containing 1× carbenicillin & 12×IPTG.Cultures were incubated with shaking ˜600 RPM 30° C. overnight.Streptavidin coated plates were bound with 50 uL of biotinylated TRCW5antigen (CD3δε-Fc-Avi, SEQ ID NO: 39) at the concentrations indicated inthe ELISA graph for 45 min at room temperature with shaking (FIG. 1 ,FIG. 3 , FIG. 4 ) followed by washing 3× with 1×TBST. Plates wereblocked with 200 ul 3% Milk in 1×TBST for ˜45 mins at room temperaturefollowed by washing 3× with 1×TBST. E. coli cultures were harvested bycentrifugation at 35000 RPMs for 10 mins 4° C. and 50 uL of supernatantwas transferred into CD3-coated plates followed by incubation at 4 C for45 min. Plates were washed 3× with 1×TBST. Bound scFv was detected withChicken Polyclonal Anti-HA-HRP (ab1190) [1:1000] for ˜45 mins RT andluminescence detected with chemi-luminescent substrate.

TRCW5 antigen (CD3δϵ-Fc-Avi, SEQ ID NO: 39)FKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMGSADDAKKDAAKKDDAKKDDAKKDGSDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVSPPSPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGLNDIFEAQKIEWHE

Clones exhibiting binding greater than the murine parent were selectedfor sequencing and exposure to titration and thermal stressing ELISAassays. Briefly, scFv were expressed as above and subjected to thermalstress (60° C. heat shock for ˜10 min) followed by ELISA analysis, asdescribed above. Briefly, the clones containing the differentcombinations of mouse back-mutations that displayed binding from E. colisupernatant were sequenced to determine which residue at each site(either human or mouse germline residue) was more optimal to maintainthermal stability. The proportion of clones harboring each residue ateach site were determined (FIGS. 2A and 2B). From the two heavy chainlibraries designed, 8 human adapted heavy chain sequences were selectedand, from the single light chain library designed, 8 human adapted lightchains were selected based on retention of >70% binding (compared toroom temperature ELISA binding) after thermal stress. Sequences of thethermally stable humanized Cris-7 VH and VL are shown below (Table 8).

TABLE 8 Sequences of thermally stable humanized Cris-7 VH and VL VHSEQ ID Variant VH Sequence NO: VDQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGL 48 000043392EWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSS VDQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGL 49 000043400EWIGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSS VDQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGL 50 000043401EWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSS VDQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGL 51 000043403EWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSS VDQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 52 000043404EWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSS VDQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 53 000043402EWIGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSS VDQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 54 000043405EWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSS VDQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 55 000043406EWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSS VL Variant VL Sequence VDEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLI 56 000043397YDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP PTFGGGTKVEIK VDEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG QSPRR LI 57 000043398YDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP PTFGGGTKVEIK VDEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 58 000043391YDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP PTFGGGTKVEIK VDEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQ SPRRWI 59 000043393YDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP PTFGGGTKVEIK VDEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGT APRR LI 60 000043394YDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP PTFGGGTKVEIK VDEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGTSPRR LIY 61 000043395DSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPP TFGGGTKVEIK VDEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGT APRRWI 62 000043396YDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP PTFGGGTKVEIK VDEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGTSPRRWI 63 000043399YDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP PTFGGGTKVEIK

The 8 new heavy chains and 8 new light chains (shown in Table 8) wereagain matrixed with each other to generate scFvs (Table 9) and furtherexposed to additional assays including titration, thermal stress, andcell binding (FIG. 3 ).

TABLE 9 Protein identities of matrixed thermally stable variable domainsVL VH VD000043397 VD000043398 VD000043391 VD000043393 VD000043394VD000043395 VD000043396 VD000043399 VD000043392 CD3B2084 CD3B2083CD3B2089 CD3B2088 CD3B2087 CD3B2086 CD3B2085 CD3B2082 VD000043400CD3B2076 CD3B2075 CD3B2081 CD3B2080 CD3B2079 CD3B2078 CD3B2077 CD3B2074VD000043401 CD3B2068 CD3B2067 CD3B2073 CD3B2072 CD3B2071 CD3B2070CD3B2069 CD3B2066 VD000043403 CD3B2057 CD3B2056 CD3B2045 CD3B2044CD3B2043 CD3B2042 CD3B2041 CD3B2055 VD000043404 CD3B2054 CD3B2053CD3B2040 CD3B2039 CD3B2038 CD3B2037 CD3B2036 CD3B2052 VD000043402CD3B2060 CD3B2059 CD3B2065 CD3B2064 CD3B2063 CD3B2062 CD3B2061 CD3B2058VD000043405 CD3B2051 CD3B2050 CD3B2035 CD3B2034 CD3B2033 CD3B2032CD3B2031 CD3B2049 VD000043406 CD3B2048 CD3B2047 CD3B2030 CD3B2029CD3B2028 CD3B2027 CD3B2026 CD3B2046

Cell binding was performed against Pan T-cells (Biological SpecialtyCorp, Item #215-02-11) and Jurkat-CD3-negative cells (ATCC® TIB-153™) toobserve an increase in non-specific binding when thermally stressed.Molecules exhibiting an increase in binding to the negative cell linewhen thermally stressed were not chosen for additional characterization.ELISA assays for binding to recombinant CD3 (TRCW5, SEQ ID NO: 39) wereperformed as above. Cell binding was performed using primary human Tcell and Jurkat cells by flow cytometry. Briefly, E. coli expressed antiCD3 ScFv supernatants, either at room temperature, or 55° C., 60° C. or65° C. heat treated samples.

Pan T-cells (CD3-positive) and Jurkat-CD3-negative cells were preparedby staining Jurkat-CD3-negative cells with CFSE while keeping panT-cells unstained. Jurkat-CD3-negative cell suspensions were resuspendedat 20 million cells per each 50 mL conical tube. Cells were harvested bycentrifugation at 400×g for 5 minutes and resuspended in DPBS followedby washing 2× in DPBS. Cells were stained with 1:25,000 dilution for afinal dye concentration of 0.02 uM CFSE (2 uL of staining solution to 50mL of cell suspension). Cells were incubated for 10 minutes at roomtemperature and centrifuged for 5 minutes at 400×g. After removal ofsupernatant, 3 mL of HI-FBS were added to the cell pellet, mixed andcentrifuged for 5 min at 400×g. Supernatant was removed and cells wereresuspended in BD stain buffer at 2×10{circumflex over ( )}6 cells/mLand incubated on ice or 4 C protected from light. Human pan T cells werethawed at 37 C and transferred gently into a conical tube containing 15mL of warm or RT culture media (RPMI+10% FBS). T-cells from Donor ID#M7348; Lot #LS 11 62980A were at 97.2% viability. Cells were harvestedby centrifugation at 400×g for 5 min and resuspended in culture media(RPMI+10% HI-FBS). Pan T-cells (without staining) were prepared at2×10{circumflex over ( )}6 cells/mL in flow staining buffer. Equalvolumes of CFSE stained Jurkat-CD3-negative cells and unstained T-cells(CD3-positive) cells were mixed and plated 50 uL/well into assay plates.50 uL/well neat bacterial supernatant samples and control sample (CD3W36ScFv) were added to each well and plates were incubated 1 hr at 4 C or.Cells were harvested by centrifugation at 400×g for 4 min and 150 uLstaining buffer were added to all wells, followed by centrifugation at400×g for 4 min to pellet cells. To each well, 150 uL staining bufferwere added followed by centrifugation at 400×g for 4 minutes to pelletcells. A647 conjugated anti-HIS secondary antibody was prepared at 2ug/mL (1:100 dilution from stock vial) in staining buffer and added at50 uL/well to the washed cells followed by incubation for 30 min at 4 Cprotected from light. Then 150 uL staining buffer were added to allwells, and plates were spun at 400×g for 4 minutes to pellet cells. 150uL IntelliCyt running buffer were added to all wells, and plates werespun at 400×g for 4 minutes to pellet cells. Cells were resuspended in20-30 uL running buffer containing 1:1000 dilution of Sytox Blue deadcell stain and run plates on iQue Screener. Briefly, cells were gated onFCS v. SCS to eliminate debris. Singlets were gated on SCS-A vs SCS-Hand, separated and gated cells first on BL1 channel based on CFSEstaining, then on VL1 channel with low/negative with Sytox blueviability stain for gating live cells. Binding of scFv molecules wasassessed by Geomeans on RL1 channel from the live cell population. Datawere analyzed in GraphPad Prism. Ultimately, four humanized matrixedclones exhibited the most desired properties: having retained bindingafter 65° C. heat shock and were selected. These clones were noted asCD3B2029, CD3B2030, CD3B2051, and CD3B2089.

Tables 10, 11, and 12 show amino acid, DNA, and CDR sequences of VH andVL for CD3B2029, CD3B2030, CD3B2051, and CD3B2089.

Table 13 shows thermal stability ELISA binding to recombinant CD3(TRCW5, SEQ ID NO: 39) data for all the clones tested from Table 9.

Table 14 shows cell stability ELISA binding data for select clones.

TABLE 10VH and VL amino acid sequences of CD3B2029, CD3B2030, CD3B2051, and CD3B2089.Binding VH VL domain SEQ ID SEQ ID name VH amino acid Sequence NO:VL amino acid sequence NO: CD3B2029 QVQLVQSGAEVKKPGSSVK 55EIVLTQSPATLSASPGERVT 59 VSCKASGYTFTRSTMHWV LSCSASSSVSYMNWYQQKKQAPGQGLEWIGYINPSSA PGQSPRRWIYDSSKLASGV YTNYNQKFQGRVTLTADKSPARFSGSGSGRDYTLTISSL TSTAYMELSSLRSEDTAVY EPEDFAVYYCQQWSRNPPYCASPQVHYDYNGFPYWG TFGGGTKVEIK QGTLVTVSS CD3B2030 QVQLVQSGAEVKKPGSSVK 55EIVLTQSPATLSASPGERVT 58 VSCKASGYTFTRSTMHWV LSCSASSSVSYMNWYQQKKQAPGQGLEWIGYINPSSA PGQAPRRWIYDSSKLASGV YTNYNQKFQGRVTLTADKSPARFSGSGSGRDYTLTISSL TSTAYMELSSLRSEDTAVY EPEDFAVYYCQQWSRNPPYCASPQVHYDYNGFPYWG TFGGGTKVEIK QGTLVTVSS CD3B2051 QVQLVQSGAEVKKPGSSVK 54EIVLTQSPATLSASPGERVT 56 VSCKASGYTFTRSTMHWV LSCSASSSVSYMNWYQQKKQAPGQGLEWMGYINPSSA PGQAPRRLIYDSSKLASGV YTNYNQKFQGRVTLTADKSPARFSGSGSGRDYTLTISSL TSTAYMELSSLRSEDTAVY EPEDFAVYYCQQWSRNPPYCASPQVHYDYNGFPYWG TFGGGTKVEIK QGTLVTVSS CD3B2089 QVQLVQSGAEVKKPGSSVK 48EIVLTQSPATLSASPGERVT 58 VSCKASGYTFTRSTMHWVR LSCSASSSVSYMNWYQQKQAPGQGLEWMGYINPSSAY PGQAPRRWIYDSSKLASGV TNYAQKFQGRVTLTADKSTPARFSGSGSGRDYTLTISSL STAYMELSSLRSEDTAVYY EPEDFAVYYCQQWSRNPPCASPQVHYDYNGFPYWGQ TFGGGTKVEIK GTLVTVSS

TABLE 11VH and VL DNA sequences of CD3B2029, CD3B2030, CD3B2051, and CD3B2089.Binding VH VL domain SEQ ID SEQ ID name VH DNA Sequence NO:VL DNA sequence NO: CD3B2029 caggttcagctggttcagtctggcgccgaa 64gagatcgtgctgacccagtctcctgccac 67 gtgaagaaacctggctcctccgtcaaggtgactgtcagcctctccaggcgagagagtca tcctgcaaggcttccggctacacctttaccaccctgtcctgctccgcttcctcctccgtgtc gatccaccatgcactgggtcaaacaggctcctacatgaactggtatcagcagaagcccg caggacaaggcttggagtggatcggctacgccagtctcctagacggtggatctacgac atcaaccccagctccgcctacaccaactactcctccaagctggcctctggcgtccctgc aaccagaaattccagggcagagtcaccctcccgcttttccggctctgggtctggcagag accgccgacaagtctacctccaccgcctacactataccctgaccatctccagcctggaa atggaactgtccagcctgagatctgaggaccctgaggacttcgccgtgtactactgcca accgccgtgtactactgcgccagccctcaggcagtggtctagaaaccctcctacctttgg gtgcactacgactacaacggcttcccttattcggaggcaccaaggtggaaatcaag ggggccagggcaccctggttaccgtttctt ct CD3B2030caggttcagctggttcagtctggcgccgaa 64 gagatcgtgctgacccagtctcctgccac 68gtgaagaaacctggctcctccgtcaaggtg actgtcagcctctccaggcgagagagtcatcctgcaaggcttccggctacacctttacca ccctgtcctgctccgcttcctcctccgtgtcgatccaccatgcactgggtcaaacaggctc ctacatgaactggtatcagcagaagcccgcaggacaaggcttggagtggatcggctac gccaggctcctagacggtggatctacgaatcaaccccagctccgcctacaccaactac ctcctccaagctggcctctggcgtccctgaaccagaaattccagggcagagtcaccctc cccgcttttccggctctggctctggcagagaccgccgacaagtctacctccaccgcctac actataccctgaccatctccagcctggaaatggaactgtccagcctgagatctgaggac cctgaggacttcgccgtgtactactgccaaccgccgtgtactactgcgccagccctcag gcagtggtctagaaaccctcctacctttgggtgcactacgactacaacggcttcccttatt cggaggcaccaaggtggaaatcaagggggccagggcaccctggttaccgtttctt ct CD3B2051caggttcagctggttcagtctggcgccgaa 65 gagatcgtgctgacccagtctcctgccac 69gtgaagaaacctggctcctccgtcaaggtg actgtcagcctctccaggcgagagagtcatcctgcaaggcttccggctacacctttacca ccctgtcctgctccgcttcctcctccgtgtcgatccaccatgcactgggtcaaacaggctc ctacatgaactggtatcagcagaagcccgcaggacaaggcttggagtggatgggctac gccaggctcctagacggctgatctacgacatcaaccccagctccgcctacaccaactac tcctccaagctggcctctggcgtccctgcaaccagaaattccagggcagagtcaccctc ccgcttttccggctctgggtctggcagagaccgccgacaagtctacctccaccgcctac actataccctgaccatctccagcctggaaatggaactgtccagcctgagatctgaggac cctgaggacttcgccgtgtactactgccaaccgccgtgtactactgcgccagccctcag gcagtggtctagaaaccctcctacctttggcggaggcaccaaggtggaaatcaag gttcactacgactacaacggcttcccttattggggccagggcaccctggttaccgtttcttct CD3B2089 caggttcagctggttcagtctggcgccgaa66 gagatcgtgctgacccagtctcctgccac 68 gtgaagaaacctggctcctccgtcaaggtgactgtcagcctctccaggcgagagagtca tcctgcaaggcttccggctacacctttaccaccctgtcctgctccgcttcctcctccgtgtc gatccaccatgcactgggtccgacaggctcctacatgaactggtatcagcagaagcccg caggccaaggcttggagtggatgggctacgccaggctcctagacggtggatctacga atcaaccccagctccgcctacaccaactacctcctccaagctggcctctggcgtccctg gcccagaaattccagggcagagtcaccctcccgcttttccggctctggctctggcagag caccgccgacaagtctacctccaccgcctaactataccctgaccatctccagcctggaa catggaactgtccagcctgagatctgaggacctgaggacttcgccgtgtactactgcca caccgccgtgtactactgcgccagccctcagcagtggtctagaaaccctcctacctttgg ggtgcactacgactacaacggcttcccttatcggaggcaccaaggtggaaatcaag tggggccagggcaccctggttaccgtttctt ct

TABLE 12CDR amino acid sequences of CD3B2029, CD3B2030, CD3B2051, and CD3B2089, usingdifferent delineations. HCDR1 HCDR2 HCDR3 (SEQ ID NO: ) (SEQ ID NO:)(SEQ ID NO: ) CD3B2029 Kabat RSTMH YINPSSAYTNYNQKFQG PQVHYDYNGFPYCD3B2089 (70) (71) (72) CD3B2030 Chothia GYTFTRS NPSSAY PQVHYDYNGFPCD3B2051 (73) (74) (75) IMGT GYTFTRST INPSSAYT ASPQVHYDYNGFPY (76) (77)(78) LCDR1 LCDR2 LCDR3 (SEQ ID NO:) (SEQ ID NO:) (SEQ ID NO:) CD3B2029Kabat SASSSVSYMN DSSKLAS QQWSRNPPT CD3B2030 (79) (80) (81) CD3B2051Chothia SSSVSY DSS WSRNPP CD3B2089 (82) (83) (84) IMGT SSVSY DSSQQWSRNPPT (85) (83) (81)

TABLE 13 Percent retained binding to recombinant CD3 after heatexposure, determined by ELISA. % Retained % Retained % Retained Clone55° C. 60° C. 65° C. CD3B2030 NtoQ 111 105 55 CD3B2040 107 106 52CD3B2065 115 112 48 CD3B2035 110 112 47 CD3B2039 111 103 44 CD3B2089 111108 41 CD3B2050 108 102 41 CD3B2081 112 114 38 CD3B2034 103 103 38CD3B2030 106 101 36 CD3B2031 108 104 36 CD3B2036 108 105 35 CD3B2064 111107 33 CD3B2051 109 94 32 CD3B2053 115 112 30 CD3B2046 111 113 30CD3B2049 108 105 29 CD3B2029 100 97 29 CD3B2026 106 98 29 CD3B2045 111105 28 CD3B2073 113 110 27 CD3B2088 116 114 26 CD3B2052 112 111 25CD3B2037 102 103 23 CD3B2054 106 100 23 CD3B2077 106 107 22 CD3B2038 10598 22 CD3B2048 105 95 22 CD3B2060 107 100 21 CD3B2058 112 110 20CD3B2032 106 88 20 CD3B2085 112 117 18 CD3B2069 114 109 17 CD3B2080 109108 17 CD3B2072 117 111 17 CD3B2041 104 97 16 CD3B2047 106 98 15CD3B2028 104 92 15 CD3B2062 110 102 14 CD3B2044 103 102 14 CD3B2059 11298 13 CD3B2066 109 109 11 CD3B2033 100 84 11 CD3B2063 108 92 11 CD3B2055102 96 11 CD3B2068 103 88 10 CD3B2082 114 116 9 CD3B2027 104 93 9CD3B2084 108 91 9 CD3B2061 116 107 9 CD3B2056 101 80 9 CD3B2076 100 82 7CD3B2070 107 82 6 CD3B2075 103 87 6 CD3B2074 104 101 6 CD3B2087 104 94 5CD3B2067 98 63 5 CD3B2086 104 96 5 CD3B2083 104 84 4 CD3B2043 100 88 4CD3B2071 103 88 4 Murine LH 10 2 3 CD3B2057 92 66 3 CD3B2079 111 92 2CDR Graft (LH) 21 3 2 CD3B2078 107 88 2 CD3B2042 96 83 2 CD3B1130 90 111 Murine HL 19 0 1 Cris7B 1 1 1

TABLE 14 shows thermal stability cell binding data. % Binding retainedon T-cells Sample ID RT 55° C. 60° C. 65° C. CD3B2030 1 1.0486771.095739 0.756759 CD3B2039 1 1.299133 1.003692 0.609466 CD3B2031 11.122011 1.204702 0.543271 CD3B2035 1 1.060978 1.141319 0.454924CD3B2041 1 1.064596 1.110962 0.44659 CD3B2034 1 0.866167 0.7837230.387208 CD3B2085 1 1.008827 0.410542 0.384652 CD3B2073 1 0.9932731.099256 0.348245 CD3B2029 1 1.058795 1.08433 0.328698 CD3B2077 11.029112 1.098231 0.306981 CD3B2045 1 1.046729 0.996753 0.29597 CD3B20331 1.072045 0.9995 0.295245 CD3B2038 1 1.027964 1.095242 0.2791 CD3B20821 1.107911 0.681799 0.275996 CD3B2088 1 1.014945 0.46063 0.257713CD3B2064 1 1.173944 0.741192 0.240813 CD3B2044 1 0.78559 1.0403650.198246 CD3B2063 1 1.004685 0.996275 0.184745 CD3B2058 1 1.0527390.807821 0.174434 CD3B2065 1 1.142664 1.33233 0.155581 CD3B2071 11.014145 0.898827 0.119915 CD3B2079 1 1.059072 0.978707 0.116397CD3B2089 1 1.084227 0.494349 0.111322 CD3B2069 1 1.076064 1.4791880.110357 CD3B2087 1 1.01658 0.963514 0.105331 CD3B2081 1 1.0351791.204711 0.10163 CD3B2072 1 0.872069 1.269992 0.093343 CD3B2042 10.992719 0.934616 0.084226 CD3B2043 1 0.990763 0.906431 0.080453CD3B2037 1 1.068237 0.986003 0.060975 CD3B2078 1 1.005626 0.7617670.057399 CD3B2080 1 1.070525 1.133895 0.05733 CD3B2086 1 1.0071521.119914 0.040405 Murine 1 0.405601 0.079161 0.051493

Mitigation of Post-Translational Modification Risks

It was determined that the parent molecule contained an “NG” motif inCDRH3 at positions #106-107, wherein the position number is counted fromthe N-terminus of the VH of CD3B2029, CD3B2030, CD3B2051, or CD3B2089sequence (SEQ ID NOs: 55, 54, or 48). The “NG” motif could potentiallypresent a risk of Post Translational Modification (PTM), specificallyAsn deamidation, and lead to loss of activity. To mitigate this PTMrisk, selected humanized variants CD3B2029, CD3B2030, CD3B2051, andCD3B2089, were further mutated at the N106 position, respectively, usingmolecular biology techniques well known in the art (Tables 15 and 16).Position N106 was mutated to one of the following residuesA/G/S/F/E/T/R/V/I/Y/L/P/Q/K. These new variants were again exposed tovarious assays including titration, thermal stress, and cell binding, asdescribed above. EC50 values for binding to Pan T-cells and Jurkatcells, as determined by ELISA are shown in Table 17, and the bindingcurves for CD3B2030 variants to recombinant CD3 (TRCW5, SEQ ID NO: 39),as an example, are shown in FIG. 4 . The % retained binding followingthe indicated heat exposure is shown in Table 18. From these assays, 4separate amino acid substitutions for the N position were selected forfurther testing. Most mutations at N106 maintained binding to somedegree, and all were considered valuable since they provided a way toboth tune the efficiency of T cell redirection and they couldsuccessfully eliminate risk of deamidation at N106.

Table 15 shows the variant CDR sequences made using CD3B2029, CD3B2030,CD3B2051, and CD3B2089 sequences.

Table 16 shows the list of substitutions in HCDR3 sequences that weremade using CD3B2029, CD3B2030, CD3B2051, and CD3B2089 sequences, whereinthe position number is counted from the N-terminus of the VH ofCD3B2029, CD3B2030, CD3B2051, or CD3B2089 (SEQ ID NOs: 55, 54, or 48).

TABLE 15CDR amino acid sequences of CD3B2029, CD3B2030, CD3B2051, and CD3B2089, usingKabat delineation. HCDR1 HCDR2 LCDR2 LCDR3 (SEQ ID (SEQ ID HCDR3 LCDR1(SEQ ID (SEQ ID NO:) NO:) (SEQ ID NO:) (SEQ ID NO:) NO:) NO:) CD3B2029RSTMH YINPSSAYT PQVHYDYXG SASSSVSYMN DSSKLAS QQWSRNPPT CD3B2030 (70)NYNQKFQG FPY, (79) (80) (81) CD3B2051 (71) wherein X CD3B2089can be Q, A, G, or S (86)

TABLE 16 Substitutions in HCDR3 sequences that were made in CD3B2029,CD3B2030, CD3B2051, and CD3B2089 sequences, wherein the position numberwas counted from the N-terminus of the VH of CD3B2029, CD3B2030,CD3B2051, or CD3B2089 (SEQ ID NOs: 55, 54, or 48). CD3 variant CDRSubstitution CD3B2029 Heavy Chain CDR3 N106Q CD3B2029 Heavy Chain CDR3N106A CD3B2029 Heavy Chain CDR3 N106G CD3B2029 Heavy Chain CDR3 N106SCD3B2030 Heavy Chain CDR3 N106Q CD3B2030 Heavy Chain CDR3 N106A CD3B2030Heavy Chain CDR3 N106G CD3B2030 Heavy Chain CDR3 N106S CD3B2051 HeavyChain CDR3 N106Q CD3B2051 Heavy Chain CDR3 N106A CD3B2051 Heavy ChainCDR3 N106G CD3B2051 Heavy Chain CDR3 N106S CD3B2089 Heavy Chain CDR3N106Q CD3B2089 Heavy Chain CDR3 N106A CD3B2089 Heavy Chain CDR3 N106GCD3B2089 Heavy Chain CDR3 N106S

TABLE 17 EC50 values for N106 PTM-mitigated, humanized Cris-7 variants.ID EC₅₀ (nM) CD3B2030 8.844 CD3B2030N106F 9.699 CD3B2030N106Q 10.04CD3B2030N106H 10.57 CD3B2030N106R 10.57 CD3B2030N106L 11.36CD3B2030N106K 11.66 CD3B2030N106A 12.77 CD3B2030N106G 14.28CD3B2030N106S 19.08 CD3B2030N106I ~13.27 CD3B2030N106P ~23.96CD3B2051N106E 8.046 CD3B2051 8.727 CD3B2051N106H 10.86 CD3B2051N106P10.89 CD3B2051N106S 11.12 CD3B2051N106R 11.2 CD3B2051N106K 11.4CD3B2051N106A 12.07 CD3B2051N106G 13.76 CD3B2051N106V 15.15 CD3B20898.794 CD3B2089N106S 10.02 CD3B2089N106F 10.13 CD3B2089N106E 10.3CD3B2089N106T 11.08 CD3B2089N106R 11.25 CD3B2089G107F 14.4 CD3B2089G107V17.39 CD3B2089N106I 17.85 CD3B2089G107Y 21.4 CD3B2089N106L 30.8CD3B2089N106P ~27.4 CD3B2089N106G ~429

TABLE 18 Thermal stability analysis of PTM-mitigated, humanized Cris-7variants. % Retained % Retained % Retained 55° C. 60° C. 65° C. CD3B2030N1060 109.6%  97.5% 72.3% CD3B2030 N106H 103.5%  98.9% 55.7% CD3B2030N106A 111.0% 102.7% 55.6% CD3B2030 N106R 107.1% 104.2% 53.8% CD3B2030104.4% 102.4% 41.3% CD3B2030 N106L 107.2% 100.5% 25.6% CD3B2030 N106F106.1%  83.3% 21.8% CD3B2030 N106K  99.7%   87.2%  7.3% CD3B2030 N106G114.7%  59.8%  1.7% CD3B2030 N106S  64.6%  19.0%  0.6% CD3B2051 N106A 98.9%  97.0% 33.3% CD3B2051 103.9%  95.0% 10.9% CD3B2051 N106H 103.1% 84.2%  9.0% CD3B2051 N106R 101.2%  74.5%  8.4% CD3B2051 N106K 116.5% 91.3%  4.0% CD3B2051 N106S 100.9%  70.9%  1.5% CD3B2051 N106G  98.9% 33.1%  1.5% CD3B2051 N106V  54.9%   1.4%  0.7% CD3B2051 N106E  78.4% 15.4%  0.5% CD3B2051 N106P  87.7%  32.6%  0.2% CD3B2089 N106R 101.6%103.0% 60.4% CD3B2089 N106F 112.7% 101.3% 36.2% CD3B2089 N106E 113.3%108.3% 33.2% CD3B2089 104.5% 109.2% 15.1% CD3B2089 N106S 102.3%  90.2% 5.4% CD3B2089 N106T 108.4%  76.7%  1.4% CD3B2089 N106I  40.6%   2.5% 0.9%

Based on the ELISA data, we chose two substitutions which had similaraffinities to the parent molecule. For the NG Motif, we chosesubstituting a Q or A for the N106 position. Additionally, it wasdesired to obtain potential substitutions which may lower the affinity.For the NG Motif, we chose G and S for the N position (which modestlylowered the affinity, based on ELISA data). These variants were thenformatted as bsAbs for further analysis for their abilities to mediatecytotoxicity and for their biophysical characteristics. Note thatCris-7-based scFv moieties were formatted in both LH and in HLorientation in the bsAbs. LH orientation provided additional ability tomodulate the affinity for CD3 and thus to tune the efficiency of T cellredirection.

Table 19 shows sequences for select CD3-specific variants.

TABLE 19Sequences for select CD3 specific variants, using Kabat delineation.. IDHeavy Chain Light Chain CD3B2030 CDR1: RSTMH (SEQ ID NO: 70)CDR1: SASSSVSYMN (SEQ ID NO: 79) CDR2: YINPSSAYTNYNQKFQG (SEQ ID NO: 71)CDR2: DSSKLAS (SEQ ID NO: 80) CDR3: PQVHYDYNGFPY (SEQ ID NO: 72)CDR3: QQWSRNPPT (SEQ ID NO: 81) VH: VL: QVQLVQSGAEVKKPGSSVKVSCKASGYEIVLTQSPATLSASPGERVTLSCSAS TFTRSTMHWVKQAPGQGLEWIGYINPSSSVSYMNWYQQKPGQAPRRWIYD SAYTNYNQKFQGRVTLTADKSTSTAYSSKLASGVPARFSGSGSGRDYTLTI MELSSLRSEDTAVYYCASPQVHYDYNGSSLEPEDFAVYYCQQWSRNPPTFG FPYWGQGTLVTVSS (SEQ ID NO: 55)GGTKVEIK (SEQ ID NO: 58) VH DNA sequence (SEQ ID NO: 64)VL DNA sequence (SEQ ID NO: 68) CD3B2030- CDR1: RSTMH (SEQ ID NO: 70)CDR1: SASSSVSYMN (SEQ ID NO: 79) N106ACDR2: YINPSSAYTNYNQKFQG (SEQ ID NO: 71) CDR2: DSSKLAS (SEQ ID NO: 80)CDR3: PQVHYDYAGFPY (SEQ ID NO: 87) CDR3: QQWSRNPPT (SEQ ID NO: 81) VH:VL: QVQLVQSGAEVKKPGSSVKVSCKASGY EIVLTQSPATLSASPGERVTLSCSASTFTRSTMHWVKQAPGQGLEWIGYINPS SSVSYMNWYQQKPGQAPRRWIYDSAYTNYNQKFQGRVTLTADKSTSTAY SSKLASGVPARFSGSGSGRDYTLTIMELSSLRSEDTAVYYCASPQVHYDYAG SSLEPEDFAVYYCQQWSRNPPTFGFPYWGQGTLVTVSS (SEQ ID NO: 88) GGTKVEIK (SEQ ID NO: 58) VH DNA sequenceVL DNA sequence CAGGTTCAACTGGTTCAGTCTGGCGC GAGATCGTGCTGACCCAGTCTCCCGAAGTGAAGAAACCTGGCTCCTCCG TGCCACACTGTCAGCCTCTCCAGTCAAGGTGTCCTGCAAGGCTFCCGGC GCGAGAGAGTCACCCTGTCCTGCTACACCTTTACCAGATCCACCATGCAC TCCGCTTCCTCCTCCGTGTCCTACTGGGTCAAGCAGGCCCCTGGACAAGG ATGAACTGGTATCAGCAGAAGCCCTTGGAGTGGATCGGCTACATCAACC CGGCCAGGCTCCTAGACGGTGGACCAGCTCCGCCTACACCAACTACAAC TCTACGACTCCTCCAAGCTGGCCTCAGAAATTCCAGGGCAGAGTGACCCT CTGGCGTCCCTGCCCGCTTTTCCGGACCGCCGACAAGTCTACCTCCACCG GCTCTGGCTCTGGCAGAGACTATCCTACATGGAACTGTCCAGCCTGAGA ACCCTGACCATCTCCAGCCTGGATCTGAGGACACCGCCGTGTACTACTG ACCTGAGGACTTCGCCGTGTACTCGCCTCTCCTCAGGTCCACTACGACTA ACTGCCAGCAGTGGTCTAGAAACCGCCGGCTTTCCTTATTGGGGCCAGG CCTCCTACCTTTGGCGGAGGCACGCACACTGGTCACCGTTTCTTCT (SEQ CAAGGTGGAAATCAAG (SEQ ID ID NO: 89) NO: 68)CD3B2089 CDR1: RSTMH (SEQ ID NO: 70) CDR1: SASSSVSYMN (SEQ ID NO: 79)CDR2: YINPSSAYTNYNQKFQG (SEQ ID NO: 71) CDR2: DSSKLAS (SEQ ID NO: 80)CDR3: PQVHYDYNGFPY (SEQ ID NO: 72) CDR3: QQWSRNPPT (SEQ ID NO: 81) VH:VL: QVQLVQSGAEVKKPGSSVKVSCKASGY EIVLTQSPATLSASPGERVTLSCSASTFTRSTMHWVRQAPGQGLEWMGYINP SSVSYMNWYQQKPGQAPRRWIYDSSAYTNYAQKFQGRVTLTADKSTSTAY SSKLASGVPARFSGSGSGRDYTLTIMELSSLRSEDTAVYYCASPQVHYDYNG SSLEPEDFAVYYCQQWSRNPPTFGFPYWGQGTLVTVSS (SEQ ID NO: 48) GGTKVEIK (SEQ ID NO: 58)VH DNA sequence (SEQ ID NO: 66) VL DNA sequence (SEQ ID NO: 68)CD3B2089- CDR1: RSTMH (SEQ ID NO: 70) CDR1: SASSSVSYMN (SEQ ID NO: 79)N106G CDR2: YINPSSAYTNYNQKFQG (SEQ ID NO: 71)CDR2: DSSKLAS (SEQ ID NO: 80) CDR3: PQVHYDYGGFPY (SEQ ID NO: 90)CDR3: QQWSRNPPT (SEQ ID NO: 81) VH: VL: QVQLVQSGAEVKKPGSSVKVSCKASGYEIVLTQSPATLSASPGERVTLSCSAS TFTRSTMHWVRQAPGQGLEWMGYINPSSVSYMNWYQQKPGQAPRRWIYD SSAYTNYAQKFQGRVTLTADKSTSTAYSSKLASGVPARFSGSGSGRDYTLTI MELSSLRSEDTAVYYCASPQVHYDYGGSSLEPEDFAVYYCQQWSRNPPTFG FPYWGQGTLVTVSS (SEQ ID NO: 242)GGTKVEIK (SEQ ID NO: 58) VH DNA sequence VL DNA sequence (SEQ ID NO: 68)CAGGTTCAACTGGTTCAGTCTGGCGC CGAAGTGAAGAAACCTGGCTCCTCCGTGAAAGTGTCCTGCAAGGCTTCCGGC TACACTTTTACCAGATCCACCATGCACTGGGTCCGACAGGCTCCAGGACAAGG CTTGGAGTGGATGGGCTACATCAACCCCAGCTCCGCCTACACCAACTACGCC CAGAAATTCCAGGGCAGAGTGACCCTGACCGCCGACAAGTCTACCTCCACCG CCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTG CGCTTCTCCTCAGGTGCACTACGACTACGGCGGCTTTCCTTATTGGGGCCAGG GCACACTGGTCACCGTTTCTTCT (SEQ ID NO: 91)

Epitope Identification

The epitope on CD3 was determined by hydrogen-deuterium exchange massspectrometry (HDX-MS). The antibody clone OKT3 was used as a control forthe HDX experiment, since its epitope on CD3ε was known from crystalstructure (PDB ID 1SY6) (Kjer-Nielsen, L. et al.; Proc Natl Acad Sci USA101, 7675-7680).

On-Exchange Experiment for HDX-MS. On-exchange reaction was initiated bymixing 10 μL of 10 μM CD3W220 (SEQ ID NO: 5), which was comprised ofCD3εγ fused with a 26-aa linker region fused onto a serum albumindomain, with or without 1.2 molar-excess of ligand and 30 μL of H₂O or adeuterated buffer (20 mM MES, pH 6.4, 150 mM NaCl in 95% D20 or 20 mMTris, pH 8.4, 150 mM NaCl in 95% D20). The reaction mixture wasincubated for 15, 50, 150, 500, or 1,500 s at 1.2° C. The on-exchangedsolution was quenched by the addition of chilled 40 μL of 8 M urea, 1 MTCEP, pH 3.0 and immediately analyzed.

CD3W220 (CD3ϵγ-HSA-6xHis) (SEQ ID NO: 92):QDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVGSADDAKKDAAKKDDAKKDDAKKDGSQSIKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDGKMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMGGGSDAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQSPFEDHVKLVNEVTEFAKTCVADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNECFLQHKDDNPNLPRLVRPEVDVMCTAFHDNEETFLKKYLYEIARRHPYFYAPELLFFAKRYKAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKCASLQKFGERAFKAWAVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDLLECADDRADLAKYICENQDSISSKLKECCEKPLLEKSHCIAEVENDEMPADLPSLAADFVESKDVCKNYAEAKDVFLGMFLYEYARRHPDYSVVLLLRLAKTYETTLEKCCAAADPHECYAKVFDEFKPLVEEPQNLIKQNCELFEQLGEYKFQNALLVRYTKKVPQVSTPTLVEVSRNLGKVGSKCCKHPEAKRMPCAEDYLSVVLNQLCVLHEKTPVSDRVTKCCTESLVNRRPCFSALEVDETYVPKEFNAETFTFHADICTLSEKERQIKKQTALVELVKHKPKATKEQLKAVMDDFAAFVEKCCKADDKETCFAEEGKKLVAASQAALGLGGGSHHH HHHHH

Procedure for HDX-MS Data Acquisition. HDX-MS sample preparation wasperformed with automated HDx system (LEAP Technologies, Morrisville,N.C.). The columns and pump were; protease, protease type XIII (proteasefrom Aspergillus saitoi, type XIII)/pepsin column (w/w, 1:1; 2.1×30 mm)(NovaBioAssays Inc., Woburn, Mass.); trap, ACQUITY UPLC BEH C18 VanGuardPre-column (2.1×5 mm) (Waters, Milford, Mass.), analytical, Accucore C18(2.1×100 mm) (Thermo Fisher Scientific, Waltham, Mass.); and LC pump,VH-P10-A (Thermo Fisher Scientific). The loading pump (from the proteasecolumn to the trap column) was set at 600 μL/min with 99% water, 1%acetonitrile, 0.1% formic acid. The gradient pump (from the trap columnto the analytical column) was set from 8% to 28% acetonitrile in 0.1%aqueous formic acid in 20 min at 100 μL/min.

MS Data Acquisition. Mass spectrometric analyses were carried out usingan LTQ™ Orbitrap Fusion Lumos mass spectrometer (Thermo FisherScientific) with the capillary temperature at 275° C., resolution150,000, and mass range (m/z) 300-1,800.

HDX-MS Data Extraction. BioPharma Finder 3.0 (Thermo Fisher Scientific)was used for the peptide identification of non-deuterated samples priorto the HDX experiments. HDExaminer version 2.5 (Sierra Analytics,Modesto, Calif.) was used to extract centroid values from the MS rawdata files for the HDX experiments.

HDX-MS Data Analysis. The extracted HDX-MS data were further analyzed inExcel. All exchange time points (at pH 6.4 or pH 8.4 at 1.2° C.) wereconverted to the equivalent time points at pH 7.4 and 23° C. (e.g., 15 sat pH 6.4 at 1.2° C. is equivalent of 0.15 s at pH 7.4 at 23° C.; Table20).

TABLE 20 HDX reaction conditions and exchange times versus exchangetimes corrected to pH 7.4 and 23° C. Time adjusted to pH 6.4 pH 8.4 pH7.4, 23° C. (s) 1.2° C. (s) 1.2° C. (s) 0.015 — — 0.05 — — 0.15 15 — 0.550 — 1.5 150 — 5 500 — 15 1,500 15 50 — 50 150 — 150 500 — 500 1,500 —1,500

Results. The antibody clone OKT3 was used as a control for the HDXexperiment, since its epitope on CD3ε was known from crystal structure(PDB ID 1SY6). Consistent with the crystal structure of OKT3 bound toCD3ε, the epitope of OKT3 was found to consist of peptides spanningresidues 29-37, 79-84 and 87-89. To determine the epitope on CD3ε boundby Cris7b, a bi-specific protein comprising Cris7b-N106Q formatted asFab (SEQ ID Nos: 93 and 94) and paired with an antigen-specific scFv-Fcarm was used. This experiment showed that Cris7 interacted with anepitope consisting of peptides spanning residues 33-37, 53-54, and79-84, which partially overlapped with that of OKT3 but also interactedwith a peptide spanning residues 53-54, which were unique compared toOKT3.

Cris7b-N106Q HC, SEQ ID NO 93:QVQLLQSAAEVKKPGESLKISCKGSGYTFTRSTMHWVRQTPGKGLEWMGYINPSSAYTNYNQKFKDQVTISADKSISTAYLQWSSLKASDTAMYYCARPQVHYDYQGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGCris7b-N106Q LC, SEQ ID NO: 94EIVLTQSPSAMSASVGDRVTITCSASSSVSYMNWYQQKPGKVPKRLIYDSSKLASGVPSRFSGSGSGTEYTLTISSLQPEDFATYYCQQWSRNPPTFGQGTMLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC

Example 2. Characterization of Novel CD3 Binders in a Bi-Specific Format

The VH/VL regions of the anti-CD3 antibodies generated in Example 1 andthe VH/VL regions of the anti-BCMA antibodies described below wereengineered into bispecific format and expressed as IgG1 (Table 21).

Engineering of CD3 scFvs for BCMAxCD3 Bispecific Generation.

CD3 VH/VL regions were engineered as scFvs in either VH-Linker-VL (alsotermed “HL”) or VL-linker-VH (also termed “LH”) orientations using thelinker of SEQ ID NO: 3 (Table 22). The VH-Linker-VL or VL-linker-VH scFvmolecules binding CD3 were further engineered into a scFv-hinge-CH2-CH3(also termed scFv-Fc) format comprising Fc silencing mutation(L234A/L235A/D265S) and the T350V/L351Y/F405A/Y407V mutations designedto promote selective heterodimerization (Table 23). The polypeptide ofSEQ ID NO: 95 was used as the constant domain hinge-CH2-CH3. DNAsequences of anti-CD3 molecules in scFv format and scFv-hinge-CH2-CH3format are shown in Table 24.

(huIgG1_Glm(17)-hinge-Fc_C220S_AAS_ZWA) SEQ ID NO: 95EPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

TABLE 21 CD3xBCMA bi-specific proteins. ID Description BC3B129 HC1:CD3B2089-N106S LH scFv; HC2: BCMB519 Fab BC3B128 HC1: CD3B2089-N106Q LHscFv; HC2: BCMB519 Fab BC3B127 HC1: CD3B2089-N106G LH scFv; HC2: BCMB519Fab BC3B126 HC1: CD3B2089-N106A LH scFv; HC2: BCMB519 Fab BC3B125 HC1:CD3B2089 LH scFv; HC2: BCMB519 Fab BC3B124 HC1: CD3B2051-N106S LH scFv;HC2: BCMB519 Fab BC3B123 HC1: CD3B2051-N106Q LH scFv; HC2: BCMB519 FabBC3B122 HC1: CD3B2051-N106G LH scFv; HC2: BCMB519 Fab BC3B121 HC1:CD3B2051-N106A LH scFv; HC2: BCMB519 Fab BC3B120 HC1: CD3B2051 LH scFv;HC2: BCMB519 Fab BC3B119 HC1: CD3B2030-N106S LH scFv; HC2: BCMB519 FabBC3B118 HC1: CD3B2030-N106G LH scFv; HC2: BCMB519 Fab BC3B117 HC1:CD3B2030-N106A LH scFv; HC2: BCMB519 Fab BC3B116 HC1: CD3B2030 LH scFv;HC2: BCMB519 Fab BC3B115 HC1: CD3B2089-N106S HL scFv; HC2: BCMB519 FabBC3B114 HC1: CD3B2089-N106Q HL scFv; HC2: BCMB519 Fab BC3B113 HC1:CD3B2089-N106G HL scFv; HC2: BCMB519 Fab BC3B112 HC1: CD3B2089-N106A HLscFv; HC2: BCMB519 Fab BC3B111 HC1: CD3B2089 HL scFv; HC2: BCMB519 FabBC3B110 HC1: CD3B2051-N106S HL scFv; HC2: BCMB519 Fab BC3B109 HC1:CD3B2051-N106Q HL scFv; HC2: BCMB519 Fab BC3B108 HC1: CD3B2051-N106G HLscFv; HC2: BCMB519 Fab BC3B107 HC1: CD3B2051-N106A HL scFv; HC2: BCMB519Fab BC3B106 HC1: CD3B2051 HL scFv; HC2: BCMB519 Fab BC3B105 HC1:CD3B2030-N106S HL scFv; HC2: BCMB519 Fab BC3B104 HC1: CD3B2030-N106G HLscFv; HC2: BCMB519 Fab BC3B103 HC1: CD3B2030-N106A HL scFv; HC2: BCMB519Fab BC3B102 HC1: CD3B2030 HL scFv; HC2: BCMB519 Fab BC3B53 HC1:CD3B2030-N106Q LH scFv; HC2: BCMB519 Fab BC3B51 HC1: CD3B2030-N106Q HLscFv; HC2: BCMB519 Fab HL-VH-Linker-VL fused to Fc; LH-VL-Linker-VHfused to Fe

TABLE 22 CD3 specific scFvs sequences. SEQ ID ID Amino acid sequence NO:CD3B2089-N106S  EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG  96 LH scFvQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYSGF PYWGQGTLVTVSS CD3B2089-N106Q EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG  97 LH scFvQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYQGF PYWGQGTLVTVSS CD3B2089-N106G EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG  98 LH scFvQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGF PYWGQGTLVTVSS CD3B2089-N106A EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG  99 LH scFvQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGF PYWGQGTLVTVSS CD3B2089 EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 100 LH scFvQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGF PYWGQGTLVTVSS CD3B2051-N106S EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 101 LH scFvQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYSGFP YWGQGTLVTVSS CD3B2051-N106Q EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 102 LH scFvQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYQGFP YWGQGTLVTVSS CD3B2051-N106G EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 103 LH scFvQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFP YWGQGTLVTVSS CD3B2051-N106A EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 104 LH scFvQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFP YWGQGTLVTVSS CD3B2051 EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 105 LH scFvQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFP YWGQGTLVTVSS CD3B2030-N106S EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 106 LH scFvQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYSGFP YWGQGTLVTVSS CD3B2030-N106G EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 107 LH scFvQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFP YWGQGTLVTVSS CD3B2030-N106A EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 108 LH scFvQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFP YWGQGTLVTVSS CD3B2030 EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 109 LH scFvQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFP YWGQGTLVTVSS CD3B2089-N106S QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVR 110 HL scFvQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYSGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2089-N106Q QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVR 111 HL scFvQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYQGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2089-N106G QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVR 112 HL scFvQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2089-N106A QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVR 113 HL scFvQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2089 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVR 114 HL scFvQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2051-N106S QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVK 115 HL scFvQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYSGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2051-N106Q QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVK 116 HL scFvQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYQGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2051-N106G QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVK 117 HL scFvQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2051-N106A QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVK 118 HL scFvQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2051 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVK 119 HL scFvQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2030-N106S QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVK 120 HL scFvQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYSGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2030-N106G QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVK 121 HL scFvQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2030-N106A QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVK 122 HL scFvQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2030 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVK 123 HL scFvQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2030-N106Q EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPG 124 LH scFvQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYQGFP YWGQGTLVTVSS CD3B2030-N106Q QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVK 125 HL scFvQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYQGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRN PPTFGGGTKVEIK CD3B2029-N106Q QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLE 126 HL scFvWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYQGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQSPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGG GTKVEIK

TABLE 23 CD3 specific scFv-Fc (scFv-hinge CH2-CH3) arms. SEQ ID AcronymAmino acid sequence NO: CD3B2089-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 127 N106S LHYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP scFv-FcPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYSGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2089-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 128 N106Q LHYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP scFv-FcPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYQGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2089-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 129 N106G LHYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP scFv-FcPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2089-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 130 N106A LHYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP scFv-FcPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2089EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 131 LH scFv-FcYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2051-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIY 132 N106S LHDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPT scFv-FcFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYSGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG CD3B2051-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIY 133 N106Q LHDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPT scFv-FcFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYQGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG CD3B2051-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIY 134 N106G LHDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPT scFv-FcFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG CD3B2051-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIY 135 N106A LHDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPT scFv-FcFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG CD3B2051EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIY 136 LH scFv-FcDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG CD3B2030-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 137 N106S LHYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP scFv-FcPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYSGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG CD3B2030-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 138 N106G LHYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP scFv-FcPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG CD3B2030-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 139 N106A LHYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP scFv-FcPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG CD3B2030EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 140 LH scFv-FcYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG CD3B2089-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGL 141 N106S HLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDT scFv-FcAVYYCASPQVHYDYSGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPG CD3B2089-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGL 142 N106Q HLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDT scFv-FcAVYYCASPQVHYDYQGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPG CD3B2089-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGL 143 N106G HLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDT scFv-FcAVYYCASPQVHYDYGGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPG CD3B2089-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGL 144 N106A HLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDT scFv-FcAVYYCASPQVHYDYAGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPG CD3B2089QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGL 145 HL scFv-FcEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPG CD3B2051-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 146 N106S HLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDT scFv-FcAVYYCASPQVHYDYSGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2051-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 147 N106Q HLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDT scFv-FcAVYYCASPQVHYDYQGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2051-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 148 N106G HLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDT scFv-FcAVYYCASPQVHYDYGGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2051-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 149 N106A HLEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDT scFv-FcAVYYCASPQVHYDYAGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2051QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 150 HL scFv-FcEWMGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRLIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2030-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 151 N106S HLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTA scFv-FcVYYCASPQVHYDYSGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2030-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 152 N106G HLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTA scFv-FcVYYCASPQVHYDYGGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2030-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 153 N106A HLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTA scFv-FcVYYCASPQVHYDYAGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2030QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 154 HL scFv-FcEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYNGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2030-EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWI 155 N106Q LHYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNP scFv-FcPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYQGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG CD3B2030-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 156 N106Q HLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTA scFv-FcVYYCASPQVHYDYQGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG CD3B2029-QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGL 157 N106Q HLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTA scFv-FcVYYCASPQVHYDYQGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQSPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG

TABLE 24DNA SEQ ID NOs for anti-CD3 scFv and scFv-hinge-CH2-CH3 (scFv-Fc)scFv-Fc DNA scFv DNA Acronym (SEQ ID NO:) (SEQ ID NO:) CD3B2089-GAGATTGTACTGACACAGTCCCCAGCAACCTTGTCCGCTTC GAGATTGTACTGACACAGTCC N106S-TCCCGGCGAAAGGGTCACTCTCTCCTGCTCCGCTAGTTCTT CCAGCAACCTTGTCCGCTTCTC LH-scFvCAGTGTCATATATGAATTGGTACCAACAAAAGCCAGGTCA CCGGCGAAAGGGTCACTCTCTGGCTCCAAGAAGATGGATTTACGATTCCTCCAAGTTGGCTT CCTGCTCCGCTAGTTCTTCAGTCTGGTGTCCCTGCACGATTTAGCGGGTCAGGGTCAGGGCG GTCATATATGAATTGGTACCACGATTACACACTCACAATTAGTAGTCTCGAACCCGAGGACT ACAAAAGCCAGGTCAGGCTCCTTGCCGTATATTACTGTCAGCAATGGAGTCGGAATCCCCCA AAGAAGATGGATTTACGATTCACTTTCGGCGGGGGAACAAAAGTAGAAATAAAAGGCGGC CTCCAAGTTGGCTTCTGGTGTCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAA CCTGCACGATTTAGCGGGTCAGAGCACCGGCGGCAGCCAAGTGCAGTTGGTCCAGTCAGG GGGTCAGGGCGCGATTACACACGCAGAAGTGAAGAAGCCCGGCTCAAGCGTCAAGGTATC CTCACAATTAGTAGTCTCGAACATGTAAGGCTTCTGGATATACTTTCACCCGAAGCACAATGC CCGAGGACTTTGCCGTATATTACTGGGTTCGCCAGGCCCCTGGACAGGGTCTTGAGTGGAT ACTGTCAGCAATGGAGTCGGAGGGGTATATCAACCCATCCTCAGCATATACTAACTATGCTC ATCCCCCAACTTTCGGCGGGGAGAAGTTTCAGGGGCGTGTCACTTTGACCGCCGATAAGTC GAACAAAAGTAGAAATAAAACACAAGCACCGCTTATATGGAACTGTCTTCATTGCGCTCTG GGCGGCTCCGAGGGCAAGAGAAGACACTGCAGTGTACTATTGCGCCAGCCCACAGGTCCA CAGCGGCAGCGGCAGCGAGACTACGACTATTCTGGATTTCCATACTGGGGGCAGGGGACC GCAAGAGCACCGGCGGCAGCTTGGTGACTGTAAGCTCTGAGCCCAAATCTAGCGACAAAA CAAGTGCAGTTGGTCCAGTCACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGG GGCGCAGAAGTGAAGAAGCCGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACA CGGCTCAAGCGTCAAGGTATCCCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT ATGTAAGGCTTCTGGATATACGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA TTTCACCCGAAGCACAATGCACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC CTGGGTTCGCCAGGCCCCTGGAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT ACAGGGTCTTGAGTGGATGGGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT GGTATATCAACCCATCCTCAGCGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC ATATACTAACTATGCTCAGAACAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA GTTTCAGGGGCGTGTCACTTTGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG GACCGCCGATAAGTCCACAAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG CACCGCTTATATGGAACTGTCTGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG TCATTGCGCTCTGAAGACACTAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC GCAGTGTACTATTGCGCCAGCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC CCACAGGTCCACTACGACTATTAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC CTGGATTTCCATACTGGGGGCGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA AGGGGACCTTGGTGACTGTAACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT GCTCTG (158) (159) CD3B2089-GAGATCGTACTGACCCAAAGTCCCGCCACTCTCTCTGCTAG GAGATCGTACTGACCCAAAGT N106Q-CCCAGGCGAGAGAGTTACCTTGTCTTGCTCTGCTAGTTCAA CCCGCCACTCTCTCTGCTAGCC LH-scFvGTGTCAGTTATATGAACTGGTATCAGCAGAAGCCAGGACA CAGGCGAGAGAGTTACCTTGTGGCACCTCGAAGATGGATATATGACTCCTCCAAACTCGCA CTTGCTCTGCTAGTTCAAGTGTTCAGGCGTACCAGCACGCTTTTCTGGGAGCGGTAGTGGTA CAGTTATATGAACTGGTATCAGGGATTATACACTCACCATCTCTAGTTTGGAACCAGAAGAT GCAGAAGCCAGGACAGGCACTTCGCTGTGTACTATTGCCAGCAGTGGAGCCGCAACCCTCC CTCGAAGATGGATATATGACTTACCTTCGGCGGTGGGACAAAGGTAGAAATAAAAGGCGG CCTCCAAACTCGCATCAGGCGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCA TACCAGCACGCTTTTCTGGGAAGAGCACCGGCGGCAGCCAAGTGCAGTTGGTTCAATCCG GCGGTAGTGGTAGGGATTATAGCGCTGAAGTGAAGAAACCTGGGTCATCTGTCAAAGTATC CACTCACCATCTCTAGTTTGGACTGTAAAGCCTCTGGGTACACTTTTACACGTAGCACCATGC ACCAGAAGATTTCGCTGTGTAACTGGGTCCGTCAAGCCCCTGGGCAAGGCCTTGAGTGGAT CTATTGCCAGCAGTGGAGCCGGGGTTATATAAACCCATCCTCCGCATACACAAATTACGCTC CAACCCTCCTACCTTCGGCGGTAAAAATTTCAAGGGCGAGTCACTCTCACTGCCGATAAATC GGGACAAAGGTAGAAATAAACACTTCAACTGCCTATATGGAGCTTAGTTCATTGCGATCAG AGGCGGCTCCGAGGGCAAGAAAGATACTGCAGTCTATTATTGTGCATCACCTCAGGTCCAT GCAGCGGCAGCGGCAGCGAGTACGACTACCAAGGGTTCCCCTACTGGGGACAGGGGACTT AGCAAGAGCACCGGCGGCAGTGGTAACTGTGTCTTCTGAGCCCAAATCTAGCGACAAAACT CCAAGTGCAGTTGGTTCAATCCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGG CGGCGCTGAAGTGAAGAAACGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACC CTGGGTCATCTGTCAAAGTATCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGG CCTGTAAAGCCTCTGGGTACATGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTG CTTTTACACGTAGCACCATGCAGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAA CTGGGTCCGTCAAGCCCCTGGGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGT GCAAGGCCTTGAGTGGATGGCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGC GTTATATAAACCCATCCTCCGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAG ATACACAAATTACGCTCAAAACCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCC ATTTCAAGGGCGAGTCACTCTCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGGGAG CACTGCCGATAAATCCACTTCAGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCA ACTGCCTATATGGAGCTTAGTTAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGA CATTGCGATCAGAAGATACTGGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTC CAGTCTATTATTGTGCATCACCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGCAA TCAGGTCCATTACGACTACCAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGT AGGGTTCCCCTACTGGGGACACTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACT GGGGACTTTGGTAACTGTGTCACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT TTCTG (160) (161) CD3B2089-GAGATCGTATTGACACAATCACCCGCCACATTGTCAGCTA GAGATCGTATTGACACAATCA N106G-GCCCCGGTGAGCGCGTCACACTTTCTTGTAGTGCATCAAG CCCGCCACATTGTCAGCTAGC LH-scFvTAGCGTTTCTTACATGAATTGGTATCAGCAGAAACCAGGA CCCGGTGAGCGCGTCACACTTCAAGCACCACGGCGATGGATATACGATTCTAGCAAACTCG TCTTGTAGTGCATCAAGTAGCCCAGTGGCGTCCCCGCTCGATTCTCCGGGTCTGGCAGTGG GTTTCTTACATGAATTGGTATCTAGAGATTATACACTCACTATCAGTTCTCTGGAACCAGAAG AGCAGAAACCAGGACAAGCAACTTCGCAGTCTATTACTGTCAACAATGGTCACGGAATCCC CCACGGCGATGGATATACGATCCCACATTCGGTGGTGGCACCAAGGTTGAAATTAAGGGCG TCTAGCAAACTCGCCAGTGGCGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGC GTCCCCGCTCGATTCTCCGGGAAGAGCACCGGCGGCAGCCAAGTTCAGCTTGTGCAGAGC TCTGGCAGTGGTAGAGATTATGGGGCAGAGGTGAAGAAACCCGGATCAAGCGTCAAAGTT ACACTCACTATCAGTTCTCTGGTCTTGTAAAGCTAGTGGATATACTTTCACACGCTCAACTAT AACCAGAAGACTTCGCAGTCTGCACTGGGTGAGACAAGCTCCTGGTCAGGGCCTGGAGTG ATTACTGTCAACAATGGTCACGATGGGGTACATAAATCCCTCCAGTGCATATACTAACTATG GGAATCCCCCCACATTCGGTGCTCAAAAGTTCCAAGGCCGCGTAACTCTCACTGCCGATAA GTGGCACCAAGGTTGAAATTAGTCCACCAGCACTGCCTACATGGAACTGTCTAGTTTGCGAT AGGGCGGCTCCGAGGGCAAGCCGAGGACACCGCCGTGTACTACTGTGCTTCACCTCAAGTA AGCAGCGGCAGCGGCAGCGACATTATGACTACGGGGGATTTCCCTACTGGGGCCAAGGTA GAGCAAGAGCACCGGCGGCACTTTGGTCACAGTCTCAAGCGAGCCCAAATCTAGCGACAA GCCAAGTTCAGCTTGTGCAGAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCA GCGGGGCAGAGGTGAAGAAAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGG CCCGGATCAAGCGTCAAAGTTACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTG TCTTGTAAAGCTAGTGGATATGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTC ACTTTCACACGCTCAACTATGCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAG ACTGGGTGAGACAAGCTCCTGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGT GTCAGGGCCTGGAGTGGATGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA GGGTACATAAATCCCTCCAGTATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCT GCATATACTAACTATGCTCAAACCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGG AGTTCCAAGGCCGCGTAACTCCAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCC TCACTGCCGATAAGTCCACCAGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCC GCACTGCCTACATGGAACTGTTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTG CTAGTTTGCGATCCGAGGACAGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCAC CCGCCGTGTACTACTGTGCTTCGCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGA ACCTCAAGTACATTATGACTACGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGA GGGGGATTTCCCTACTGGGGCACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC CAAGGTACTTTGGTCACAGTCCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT TCAAGCG (162) (163) CD3B2089-GAAATAGTGCTGACCCAGAGCCCCGCTACCCTTTCTGCAA GAAATAGTGCTGACCCAGAGC N106A-GTCCTGGGGAACGTGTTACATTGTCTTGTAGCGCTTCTTCA CCCGCTACCCTTTCTGCAAGTC LH-scFvTCAGTCTCCTATATGAATTGGTATCAACAAAAACCAGGACA CTGGGGAACGTGTTACATTGTAGCTCCTCGGCGGTGGATCTACGACAGTTCCAAACTTGCCT CTTGTAGCGCTTCTTCATCAGTCTGGTGTGCCTGCTCGGTTTAGTGGGTCTGGAAGTGGACG CTCCTATATGAATTGGTATCAAAGATTATACTCTGACCATCAGTTCCTTGGAACCCGAGGATT CAAAAACCAGGACAAGCTCCTTTGCTGTTTATTACTGCCAACAATGGAGTAGAAACCCTCCA CGGCGGTGGATCTACGACAGTACCTTTGGAGGTGGAACTAAGGTCGAGATAAAGGGCGGC TCCAAACTTGCCTCTGGTGTGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAA CTGCTCGGTTTAGTGGGTCTGGAGCACCGGCGGCAGCCAAGTGCAATTGGTCCAAAGTGG GAAGTGGACGAGATTATACTCAGCTGAAGTAAAAAAACCCGGCTCCTCTGTGAAGGTCAGT TGACCATCAGTTCCTTGGAACCTGCAAAGCCTCAGGGTACACCTTTACTAGGTCAACAATGC CGAGGATTTTGCTGTTTATTACACTGGGTGCGACAAGCTCCCGGTCAGGGTTTGGAGTGGA TGCCAACAATGGAGTAGAAACTGGGATACATAAACCCCTCATCAGCCTACACAAATTATGCA CCTCCAACCTTTGGAGGTGGACAAAAATTTCAGGGTCGGGTTACACTCACCGCCGACAAAT ACTAAGGTCGAGATAAAGGGCCACTTCCACTGCTTATATGGAACTTTCCTCTCTCCGCAGTG CGGCTCCGAGGGCAAGAGCAAGGACACAGCAGTGTACTATTGTGCCTCCCCTCAAGTGCAT GCGGCAGCGGCAGCGAGAGCTATGACTACGCTGGTTTCCCTTACTGGGGACAAGGTACTCT AAGAGCACCGGCGGCAGCCAGGTTACAGTTTCTTCCGAGCCCAAATCTAGCGACAAAACTC AGTGCAATTGGTCCAAAGTGGACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGG AGCTGAAGTAAAAAAACCCGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACC CTCCTCTGTGAAGGTCAGTTGCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGG CAAAGCCTCAGGGTACACCTTTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTG TACTAGGTCAACAATGCACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAA GGTGCGACAAGCTCCCGGTCAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGT GGGTTTGGAGTGGATGGGATCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGC ACATAAACCCCTCATCAGCCTAAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAG CACAAATTATGCACAAAAATTTCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCC CAGGGTCGGGTTACACTCACCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGGGAG GCCGACAAATCCACTTCCACTGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCA GCTTATATGGAACTTTCCTCTCAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGA TCCGCAGTGAGGACACAGCAGGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTC TGTACTATTGTGCCTCCCCTCACCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGCAA AGTGCATTATGACTACGCTGGGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGT TTTCCCTTACTGGGGACAAGGCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACT TACTCTGGTTACAGTTTCTTCCACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT G (164) (165) CD3B2089-GAAATCGTTCTCACACAGAGCCCTGCAACATTGTCAGCTTC GAAATCGTTCTCACACAGAGC LH-scFvACCCGGTGAACGAGTAACATTGTCCTGTTCTGCCTCAAGTA CCTGCAACATTGTCAGCTTCACGTGTGAGCTATATGAATTGGTATCAACAAAAACCAGGGCA CCGGTGAACGAGTAACATTGTGGCCCCTAGAAGGTGGATCTATGATTCAAGCAAACTGGCA CCTGTTCTGCCTCAAGTAGTGTTCCGGCGTCCCTGCCCGCTTTAGTGGAAGCGGTTCAGGAA GAGCTATATGAATTGGTATCAGGGACTATACTCTTACTATCTCCAGCCTTGAACCTGAAGAT ACAAAAACCAGGGCAGGCCCCTTTGCAGTCTACTACTGCCAACAATGGTCTAGGAATCCCCC TAGAAGGTGGATCTATGATTCCACTTTTGGTGGAGGGACCAAAGTTGAGATCAAAGGCGG AAGCAAACTGGCATCCGGCGTCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCA CCCTGCCCGCTTTAGTGGAAGAGAGCACCGGCGGCAGCCAGGTACAACTCGTGCAAAGTG CGGTTCAGGAAGGGACTATACGTGCTGAAGTGAAGAAACCTGGATCAAGCGTCAAGGTATC TCTTACTATCTCCAGCCTTGAACTGTAAAGCATCAGGATACACCTTCACACGCAGTACTATGC CCTGAAGATTTTGCAGTCTACTATTGGGTGCGTCAAGCCCCCGGACAGGGCCTGGAATGGA ACTGCCAACAATGGTCTAGGATGGGCTACATAAACCCTTCTTCCGCCTACACCAATTATGCC ATCCCCCCACTTTTGGTGGAGCAAAAGTTCCAGGGAAGGGTGACTCTGACTGCTGATAAAA GGACCAAAGTTGAGATCAAAGGTACTAGCACCGCATACATGGAACTGTCTTCACTGAGAAG GCGGCTCCGAGGGCAAGAGCCGAGGACACCGCCGTCTATTATTGTGCATCCCCCCAAGTCC AGCGGCAGCGGCAGCGAGAGACTATGATTACAACGGATTTCCTTACTGGGGCCAGGGAAC CAAGAGCACCGGCGGCAGCCCTTGGTCACCGTGTCTTCCGAGCCCAAATCTAGCGACAAAA AGGTACAACTCGTGCAAAGTGCTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGG GTGCTGAAGTGAAGAAACCTGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACA GATCAAGCGTCAAGGTATCCTCCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT GTAAAGCATCAGGATACACCTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA TCACACGCAGTACTATGCATTCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC GGGTGCGTCAAGCCCCCGGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT AGGGCCTGGAATGGATGGGCGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT TACATAAACCCTTCTTCCGCCTGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC ACACCAATTATGCCCAAAAGTCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA TCCAGGGAAGGGTGACTCTGAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG CTGCTGATAAAAGTACTAGCAGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG CCGCATACATGGAACTGTCTTCGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG ACTGAGAAGCGAGGACACCGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC CCGTCTATTATTGTGCATCCCCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC CCAAGTCCACTATGATTACAACAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC GGATTTCCTTACTGGGGCCAGGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GGAACCTTGGTCACCGTGTCTCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT TCCG (166) (167) CD3B2051-GAGATTGTACTCACCCAGTCTCCAGCTACCCTTAGTGCTTC GAGATTGTACTCACCCAGTCTC N106S-ACCTGGTGAGCGCGTGACATTGTCCTGCTCCGCAAGCTCC CAGCTACCCTTAGTGCTTCACC LH-scFvAGTGTTTCATATATGAATTGGTACCAACAAAAGCCTGGGC TGGTGAGCGCGTGACATTGTCAAGCACCACGCCGGCTGATCTACGACAGCTCCAAGCTCGC CTGCTCCGCAAGCTCCAGTGTTAAGCGGTGTACCTGCTCGCTTTTCCGGCAGCGGGTCAGGT TCATATATGAATTGGTACCAACCGAGATTATACTCTGACCATTTCATCACTCGAACCCGAAGA AAAAGCCTGGGCAAGCACCACCTTTGCAGTGTATTACTGTCAACAGTGGAGTAGGAATCCA GCCGGCTGATCTACGACAGCTCCAACATTTGGGGGTGGCACCAAGGTTGAGATAAAGGGC CCAAGCTCGCAAGCGGTGTACGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAG CTGCTCGCTTTTCCGGCAGCGCAAGAGCACCGGCGGCAGCCAAGTGCAACTCGTACAATCT GGTCAGGTCGAGATTATACTCGGCGCTGAGGTTAAGAAACCTGGTAGCTCTGTTAAAGTGT TGACCATTTCATCACTCGAACCCTTGTAAAGCATCCGGGTATACTTTTACCCGGTCAACTATG CGAAGACTTTGCAGTGTATTACACTGGGTAAAACAAGCTCCTGGACAAGGTTTGGAGTGG CTGTCAACAGTGGAGTAGGAAATGGGTTATATAAATCCCTCCTCAGCATACACTAACTACAA TCCACCAACATTTGGGGGTGGCCAGAAGTTCCAGGGGCGCGTTACCCTGACTGCCGATAAG CACCAAGGTTGAGATAAAGGAGTACTTCAACTGCTTATATGGAGCTGTCATCCCTGCGTAG GCGGCTCCGAGGGCAAGAGCCGAGGACACAGCAGTATACTACTGCGCCAGTCCACAGGTA AGCGGCAGCGGCAGCGAGAGCACTACGATTACAGTGGCTTTCCATACTGGGGGCAGGGCA CAAGAGCACCGGCGGCAGCCCTCTGGTAACAGTATCTAGTGAGCCCAAATCTAGCGACAA AAGTGCAACTCGTACAATCTGAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCA GCGCTGAGGTTAAGAAACCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGG GTAGCTCTGTTAAAGTGTCTTGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTG TAAAGCATCCGGGTATACTTTTGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTC ACCCGGTCAACTATGCACTGGAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAG GTAAAACAAGCTCCTGGACAAACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGT GGTTTGGAGTGGATGGGTTATGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA ATAAATCCCTCCTCAGCATACAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCT CTAACTACAACCAGAAGTTCCCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGG AGGGGCGCGTTACCCTGACTGCAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCC CCGATAAGAGTACTTCAACTGGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCC CTTATATGGAGCTGTCATCCCTTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTG GCGTAGCGAGGACACAGCAGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCAC TATACTACTGCGCCAGTCCACAGCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGA GGTACACTACGATTACAGTGGGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGA CTTTCCATACTGGGGGCAGGGACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC CACTCTGGTAACAGTATCTAGTCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT G (168) (169) CD3B2051-GAGATCGTGTTGACTCAAAGTCCTGCAACCCTGTCTGCTAG GAGATCGTGTTGACTCAAAGT N106Q-TCCAGGGGAGAGGGTTACTCTCAGTTGTTCTGCAAGCAGT CCTGCAACCCTGTCTGCTAGTC LH-scFvAGCGTATCCTACATGAACTGGTATCAACAAAAGCCTGGTC CAGGGGAGAGGGTTACTCTCAAGGCACCACGGCGGTTGATATATGACTCCTCCAAGTTGGC GTTGTTCTGCAAGCAGTAGCGCTCTGGGGTGCCCGCAAGATTCTCCGGGTCCGGCTCTGGC TATCCTACATGAACTGGTATCACGCGATTACACACTGACTATAAGCAGTCTGGAACCAGAGG ACAAAAGCCTGGTCAGGCACCATTTTGCCGTTTACTACTGCCAACAATGGAGCCGAAACCCC ACGGCGGTTGATATATGACTCCCAACCTTTGGAGGTGGCACTAAGGTAGAGATAAAGGGC CTCCAAGTTGGCCTCTGGGGTGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAG GCCCGCAAGATTCTCCGGGTCCAAGAGCACCGGCGGCAGCCAGGTACAGTTGGTCCAAAG CGGCTCTGGCCGCGATTACACTGGCGCAGAGGTAAAGAAACCAGGTTCTTCAGTCAAGGTA ACTGACTATAAGCAGTCTGGAAGTTGCAAGGCATCTGGATATACATTTACCCGCAGTACTAT ACCAGAGGATTTTGCCGTTTAGCATTGGGTCAAACAGGCTCCAGGACAGGGGCTTGAATG CTACTGCCAACAATGGAGCCGGATGGGTTACATCAACCCATCTAGTGCCTATACAAACTATA AAACCCCCCAACCTTTGGAGGATCAGAAATTTCAGGGCAGAGTGACTCTGACAGCCGACAA TGGCACTAAGGTAGAGATAAAATCAACCTCTACAGCATATATGGAGTTGTCCTCTCTCCGTA GGGCGGCTCCGAGGGCAAGAGTGAAGATACTGCCGTCTACTATTGTGCAAGCCCCCAAGTC GCAGCGGCAGCGGCAGCGAGCACTATGATTATCAGGGTTTCCCTTACTGGGGGCAGGGTA AGCAAGAGCACCGGCGGCAGCTTTGGTTACCGTTTCATCCGAGCCCAAATCTAGCGACAAA CCAGGTACAGTTGGTCCAAAGACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG TGGCGCAGAGGTAAAGAAACGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC CAGGTTCTTCAGTCAAGGTAAACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT GTTGCAAGGCATCTGGATATAGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA CATTTACCCGCAGTACTATGCACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC TTGGGTCAAACAGGCTCCAGGAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT ACAGGGGCTTGAATGGATGGGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT GTTACATCAACCCATCTAGTGCGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC CTATACAAACTATAATCAGAACAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA ATTTCAGGGCAGAGTGACTCTGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG GACAGCCGACAAATCAACCTCGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG TACAGCATATATGGAGTTGTCGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG CTCTCTCCGTAGTGAAGATACTAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC GCCGTCTACTATTGTGCAAGCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC CCCCAAGTCCACTATGATTATCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC AGGGTTTCCCTTACTGGGGGCGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA AGGGTACTTTGGTTACCGTTTCCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT ATCCG (170) (171) CD3B2051-GAAATTGTTCTTACACAAAGTCCTGCTACACTGTCAGCCAG GAAATTGTTCTTACACAAAGTC N106G-CCCCGGTGAGCGAGTCACATTGTCATGCTCTGCTTCCAGTA CTGCTACACTGTCAGCCAGCC LH-scFvGTGTGAGCTACATGAACTGGTACCAACAGAAACCTGGTCA CCGGTGAGCGAGTCACATTGTGGCTCCAAGGCGCTTGATATACGACAGCAGCAAACTGGCA CATGCTCTGCTTCCAGTAGTGTAGTGGTGTACCTGCTCGGTTTTCTGGATCAGGCTCAGGTA GAGCTACATGAACTGGTACCAGAGACTATACTCTCACCATTTCCTCTCTGGAACCTGAGGAC ACAGAAACCTGGTCAGGCTCCTTTGCTGTTTATTATTGCCAGCAGTGGAGTCGCAACCCTCC AAGGCGCTTGATATACGACAGCACCTTCGGTGGAGGGACAAAAGTAGAAATAAAGGGCGG CAGCAAACTGGCAAGTGGTGTCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCA ACCTGCTCGGTTTTCTGGATCAAGAGCACCGGCGGCAGCCAAGTTCAACTGGTCCAAAGCG GGCTCAGGTAGAGACTATACTGTGCTGAAGTTAAAAAGCCAGGAAGCAGTGTTAAAGTCTC CTCACCATTTCCTCTCTGGAACATGTAAGGCCAGCGGTTACACTTTTACTAGGAGTACCATG CTGAGGACTTTGCTGTTTATTACACTGGGTGAAGCAGGCCCCCGGTCAGGGTCTTGAGTGG TTGCCAGCAGTGGAGTCGCAAATGGGATATATAAACCCATCATCCGCCTACACTAATTACAA CCCTCCCACCTTCGGTGGAGGCCAAAAGTTTCAGGGTCGCGTGACTTTGACCGCCGACAAA GACAAAAGTAGAAATAAAGGTCTACCAGCACAGCCTACATGGAACTCAGTTCTCTCCGATC GCGGCTCCGAGGGCAAGAGCCGAAGATACCGCTGTATATTACTGTGCTTCCCCACAAGTAC AGCGGCAGCGGCAGCGAGAGACTATGATTACGGGGGCTTCCCATACTGGGGCCAGGGAAC CAAGAGCACCGGCGGCAGCCTCTCGTCACAGTATCATCCGAGCCCAAATCTAGCGACAAAA AAGTTCAACTGGTCCAAAGCGCTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGG GTGCTGAAGTTAAAAAGCCAGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACA GAAGCAGTGTTAAAGTCTCATCCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT GTAAGGCCAGCGGTTACACTTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA TTACTAGGAGTACCATGCACTCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC GGGTGAAGCAGGCCCCCGGTAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT CAGGGTCTTGAGTGGATGGGGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT ATATATAAACCCATCATCCGCCGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC TACACTAATTACAACCAAAAGCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA TTTCAGGGTCGCGTGACTTTGGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG ACCGCCGACAAATCTACCAGCGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG ACAGCCTACATGGAACTCAGTGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG TCTCTCCGATCCGAAGATACCAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC GCTGTATATTACTGTGCTTCCCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC CACAAGTACACTATGATTACGAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC GGGGCTTCCCATACTGGGGCCGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA AGGGAACTCTCGTCACAGTATCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT CATCCG (172) (173) CD3B2051-GAAATTGTATTGACTCAGTCCCCAGCTACATTGAGCGCAA GAAATTGTATTGACTCAGTCCC N106A-GTCCTGGCGAGAGAGTAACCCTGTCTTGTTCTGCCAGTAG CAGCTACATTGAGCGCAAGTC LH-scFvTAGTGTAAGCTACATGAACTGGTATCAGCAGAAACCCGGA CTGGCGAGAGAGTAACCCTGTCAGGCCCCACGCCGACTTATCTATGATTCAAGTAAGCTCGC CTTGTTCTGCCAGTAGTAGTGTTAGTGGGGTTCCAGCCAGATTTAGTGGTTCTGGCTCTGGA AAGCTACATGAACTGGTATCACGCGATTACACTCTGACCATTTCTTCTCTGGAGCCTGAGGA GCAGAAACCCGGACAGGCCCCCTTCGCAGTATATTACTGCCAACAATGGTCACGCAATCCAC ACGCCGACTTATCTATGATTCACAACATTCGGTGGAGGGACAAAAGTGGAAATCAAAGGCG AGTAAGCTCGCTAGTGGGGTTGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGC CCAGCCAGATTTAGTGGTTCTAAGAGCACCGGCGGCAGCCAAGTTCAGTTGGTTCAATCCG GGCTCTGGACGCGATTACACTGCGCTGAGGTCAAAAAACCTGGATCATCTGTGAAAGTCTC CTGACCATTTCTTCTCTGGAGCATGTAAGGCATCTGGTTATACCTTCACTCGGAGTACCATGC CTGAGGACTTCGCAGTATATTATTGGGTTAAGCAGGCCCCCGGTCAGGGGTTGGAGTGGA ACTGCCAACAATGGTCACGCATGGGTTACATCAACCCTTCCTCAGCCTACACAAATTATAAT ATCCACCAACATTCGGTGGAGCAGAAATTTCAGGGGCGCGTTACTCTCACCGCTGACAAGT GGACAAAAGTGGAAATCAAACCACCTCCACAGCCTATATGGAGCTGTCAAGCCTGCGGAG GGCGGCTCCGAGGGCAAGAGTGAGGATACAGCCGTATATTACTGTGCCAGTCCTCAGGTTC CAGCGGCAGCGGCAGCGAGAATTATGATTACGCTGGCTTCCCATATTGGGGTCAGGGGAC GCAAGAGCACCGGCGGCAGCTCTCGTCACTGTGTCCAGCGAGCCCAAATCTAGCGACAAA CAAGTTCAGTTGGTTCAATCCACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG GGCGCTGAGGTCAAAAAACCTGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC GGATCATCTGTGAAAGTCTCAACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT TGTAAGGCATCTGGTTATACCTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA TCACTCGGAGTACCATGCATTCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC GGGTTAAGCAGGCCCCCGGTCAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT AGGGGTTGGAGTGGATGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT TACATCAACCCTTCCTCAGCCTGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC ACACAAATTATAATCAGAAATCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA TTCAGGGGCGCGTTACTCTCAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG CCGCTGACAAGTCCACCTCCAGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG CAGCCTATATGGAGCTGTCAAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG GCCTGCGGAGTGAGGATACAAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC GCCGTATATTACTGTGCCAGTCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC CTCAGGTTCATTATGATTACGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC TGGCTTCCCATATTGGGGTCAGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GGGGACTCTCGTCACTGTGTCCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT CAGCG (174) (175) CD3B2051-GAGATAGTTCTTACACAGAGCCCTGCAACCTTGAGTGCAA GAGATAGTTCTTACACAGAGC LH-scFvGTCCAGGGGAACGGGTGACTCTGAGTTGTAGTGCTTCTAG CCTGCAACCTTGAGTGCAAGTTTCCGTAAGTTATATGAACTGGTACCAACAGAAGCCAGGT CCAGGGGAACGGGTGACTCTCAAGCACCAAGACGCCTTATCTACGACTCATCTAAACTTGC GAGTTGTAGTGCTTCTAGTTCCTAGTGGAGTGCCAGCCAGATTTTCCGGTTCAGGAAGTGGG GTAAGTTATATGAACTGGTACAGGGACTACACACTTACCATCTCATCCCTTGAGCCCGAAGA CAACAGAAGCCAGGTCAAGCATTTCGCCGTATATTACTGTCAACAATGGTCAAGAAATCCTC CCAAGACGCCTTATCTACGACTCTACATTTGGTGGTGGTACAAAAGTAGAGATCAAGGGCG CATCTAAACTTGCTAGTGGAGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGC TGCCAGCCAGATTTTCCGGTTCAAGAGCACCGGCGGCAGCCAAGTGCAGTTGGTGCAGAGT AGGAAGTGGGAGGGACTACAGGGGCTGAGGTTAAAAAGCCTGGTTCCAGTGTGAAAGTC CACTTACCATCTCATCCCTTGAAGTTGTAAAGCCTCCGGGTACACTTTTACTAGGTCAACAAT GCCCGAAGATTTCGCCGTATAGCACTGGGTCAAGCAAGCCCCCGGCCAAGGCTTGGAATG TTACTGTCAACAATGGTCAAGGATGGGGTACATAAATCCAAGCAGTGCCTACACCAACTAT AAATCCTCCTACATTTGGTGGTAACCAAAAATTTCAAGGTAGAGTAACATTGACTGCTGACA GGTACAAAAGTAGAGATCAAAGTCCACATCAACTGCTTATATGGAGCTGTCCTCTCTTCGG GGGCGGCTCCGAGGGCAAGATCTGAAGATACCGCCGTATACTATTGCGCCTCCCCCCAAGT GCAGCGGCAGCGGCAGCGAGCCACTACGACTATAACGGATTTCCCTACTGGGGACAAGGA AGCAAGAGCACCGGCGGCAGACCCTGGTAACAGTTTCTTCAGAGCCCAAATCTAGCGACAA CCAAGTGCAGTTGGTGCAGAGAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCA TGGGGCTGAGGTTAAAAAGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGG TGGTTCCAGTGTGAAAGTCAGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTG TTGTAAAGCCTCCGGGTACACGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTC TTTTACTAGGTCAACAATGCACAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAG TGGGTCAAGCAAGCCCCCGGCACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGT CAAGGCTTGGAATGGATGGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA GTACATAAATCCAAGCAGTGCATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCT CTACACCAACTATAACCAAAACCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGG ATTTCAAGGTAGAGTAACATTCAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCC GACTGCTGACAAGTCCACATCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCC AACTGCTTATATGGAGCTGTCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTG CTCTCTTCGGTCTGAAGATACCGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCAC GCCGTATACTATTGCGCCTCCCGCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGA CCCAAGTCCACTACGACTATAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGA ACGGATTTCCCTACTGGGGACACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC AAGGAACCCTGGTAACAGTTTCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT CTTCAG (176) (177) CD3B2030-GAAATTGTCCTGACTCAGTCTCCAGCCACACTGAGTGCATC GAAATTGTCCTGACTCAGTCTC N106S-TCCCGGCGAGCGGGTCACTCTTAGTTGCAGCGCCAGTTCT CAGCCACACTGAGTGCATCTC LH-scFvAGTGTATCATATATGAACTGGTATCAGCAAAAGCCAGGTC CCGGCGAGCGGGTCACTCTTAAAGCTCCCAGGCGATGGATATACGACTCATCAAAACTCGC GTTGCAGCGCCAGTTCTAGTGCTCTGGCGTCCCAGCCCGGTTCTCCGGTTCCGGCTCTGGGC TATCATATATGAACTGGTATCAGCGACTATACCCTTACAATTTCTAGCCTCGAACCAGAAGAT GCAAAAGCCAGGTCAAGCTCCTTTGCTGTATATTATTGTCAACAGTGGTCACGTAACCCACC CAGGCGATGGATATACGACTCAACCTTCGGTGGAGGGACAAAGGTCGAGATAAAAGGCGG ATCAAAACTCGCCTCTGGCGTCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCA CCCAGCCCGGTTCTCCGGTTCCAGAGCACCGGCGGCAGCCAAGTACAGCTCGTTCAGTCCG GGCTCTGGGCGCGACTATACCGTGCAGAAGTCAAGAAACCAGGAAGTAGCGTAAAAGTGT CTTACAATTTCTAGCCTCGAACCATGTAAAGCAAGTGGTTATACCTTTACACGCTCAACTATG CAGAAGATTTTGCTGTATATTACATTGGGTTAAGCAGGCTCCAGGACAAGGGCTTGAGTGG TTGTCAACAGTGGTCACGTAAATAGGATACATCAATCCATCTAGCGCCTACACAAATTATAA CCCACCAACCTTCGGTGGAGGCCAGAAGTTCCAGGGGAGAGTTACCCTCACTGCCGATAAG GACAAAGGTCGAGATAAAAGTCCACATCAACCGCCTATATGGAATTGAGTTCCCTTCGTAG GCGGCTCCGAGGGCAAGAGCTGAGGACACTGCCGTCTACTACTGTGCCTCCCCTCAGGTTC AGCGGCAGCGGCAGCGAGAGATTATGATTACTCAGGTTTTCCATACTGGGGCCAGGGCACC CAAGAGCACCGGCGGCAGCCCTCGTAACAGTAAGCAGCGAGCCCAAATCTAGCGACAAAA AAGTACAGCTCGTTCAGTCCGCTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGG GTGCAGAAGTCAAGAAACCAGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACA GGAAGTAGCGTAAAAGTGTCACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT TGTAAAGCAAGTGGTTATACCGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA TTTACACGCTCAACTATGCATTCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC GGGTTAAGCAGGCTCCAGGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT AAGGGCTTGAGTGGATAGGAGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT TACATCAATCCATCTAGCGCCTGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC ACACAAATTATAACCAGAAGTCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA TCCAGGGGAGAGTTACCCTCAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG CTGCCGATAAGTCCACATCAAGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG CCGCCTATATGGAATTGAGTTGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG CCCTTCGTAGTGAGGACACTGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC CCGTCTACTACTGTGCCTCCCCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC TCAGGTTCATTATGATTACTCAAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC GGTTTTCCATACTGGGGCCAGGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GGCACCCTCGTAACAGTAAGCCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AGCG (178) (179) CD3B2030-GAAATTGTACTCACACAAAGTCCTGCAACTTTGTCTGCCTC GAAATTGTACTCACACAAAGT N106G-ACCAGGGGAAAGAGTAACTCTTAGTTGTAGTGCTAGTTCA CCTGCAACTTTGTCTGCCTCAC LH-scFvTCCGTTTCTTATATGAATTGGTATCAGCAGAAACCCGGACA CAGGGGAAAGAGTAACTCTTAAGCACCCCGGCGGTGGATATACGATTCCAGTAAACTTGCA GTTGTAGTGCTAGTTCATCCGTAGCGGAGTCCCCGCACGTTTCAGCGGCAGTGGCTCAGGCC TTCTTATATGAATTGGTATCAGGGGACTATACCCTGACTATTTCCTCCTTGGAACCTGAGGAT CAGAAACCCGGACAAGCACCCTTTGCTGTGTACTACTGTCAGCAATGGAGTAGAAATCCTCC CGGCGGTGGATATACGATTCCCACCTTTGGAGGTGGCACTAAAGTAGAGATCAAAGGCGG AGTAAACTTGCAAGCGGAGTCCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCA CCCGCACGTTTCAGCGGCAGTAGAGCACCGGCGGCAGCCAGGTGCAACTGGTACAAAGTG GGCTCAGGCCGGGACTATACCGTGCCGAGGTGAAGAAGCCAGGGTCCAGTGTGAAAGTAT CTGACTATTTCCTCCTTGGAACCATGTAAAGCCAGCGGGTACACATTCACTAGGAGCACTAT CTGAGGATTTTGCTGTGTACTAGCACTGGGTAAAGCAAGCCCCAGGGCAAGGTTTGGAGTG CTGTCAGCAATGGAGTAGAAAGATCGGTTATATTAACCCTTCATCTGCTTATACAAATTACAA TCCTCCCACCTTTGGAGGTGGTCAGAAATTCCAAGGGAGGGTCACTTTGACCGCTGACAAG CACTAAAGTAGAGATCAAAGGTCTACCTCTACTGCATACATGGAACTCTCCAGCCTTCGTTCA CGGCTCCGAGGGCAAGAGCAGAAGACACAGCCGTTTATTACTGTGCCTCCCCACAGGTACA GCGGCAGCGGCAGCGAGAGCCTACGACTACGGTGGATTCCCATATTGGGGTCAAGGCACC AAGAGCACCGGCGGCAGCCACTTGTAACAGTATCAAGCGAGCCCAAATCTAGCGACAAAA GGTGCAACTGGTACAAAGTGGCTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGG TGCCGAGGTGAAGAAGCCAGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACA GGTCCAGTGTGAAAGTATCATCCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT GTAAAGCCAGCGGGTACACATGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA TCACTAGGAGCACTATGCACTCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC GGGTAAAGCAAGCCCCAGGGAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT CAAGGTTTGGAGTGGATCGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT TATATTAACCCTTCATCTGCTTGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC ATACAAATTACAATCAGAAATCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA TCCAAGGGAGGGTCACTTTGAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG CCGCTGACAAGTCTACCTCTACGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG TGCATACATGGAACTCTCCAGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG CCTTCGTTCAGAAGACACAGCAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC CGTTTATTACTGTGCCTCCCCACTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC CAGGTACACTACGACTACGGTAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC GGATTCCCATATTGGGGTCAAGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GGCACCCTTGTAACAGTATCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AGCG (180) (181) CD3B2030-GAAATTGTTTTGACCCAATCACCTGCCACTCTCTCTGCCTCT GAAATTGTTTTGACCCAATCAC N106A-CCTGGTGAGCGAGTTACTTTGTCATGTAGCGCATCATCAA CTGCCACTCTCTCTGCCTCTCC LH-scFvGTGTATCTTACATGAACTGGTACCAACAAAAACCCGGACA TGGTGAGCGAGTTACTTTGTCGGCACCACGTCGTTGGATTTATGACAGTAGCAAGCTCGCC ATGTAGCGCATCATCAAGTGTTCCGGGGTACCCGCAAGATTTTCCGGGTCAGGGTCTGGCA ATCTTACATGAACTGGTACCAGGGACTATACCCTGACAATCAGCAGTCTGGAACCTGAGGA ACAAAAACCCGGACAGGCACCCTTTGCTGTGTATTACTGCCAACAGTGGTCTCGCAACCCCC ACGTCGTTGGATTTATGACAGCTACTTTCGGGGGAGGTACAAAGGTAGAAATTAAGGGCG TAGCAAGCTCGCCTCCGGGGTGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGC ACCCGCAAGATTTTCCGGGTCAAGAGCACCGGCGGCAGCCAAGTGCAACTCGTGCAAAGC AGGGTCTGGCAGGGACTATACGGGGCTGAAGTGAAGAAGCCTGGATCAAGCGTGAAGGTC CCTGACAATCAGCAGTCTGGAAGTTGCAAAGCCTCTGGATATACCTTCACTCGATCAACCAT ACCTGAGGACTTTGCTGTGTAGCACTGGGTCAAGCAGGCCCCAGGGCAAGGGCTCGAATG TTACTGCCAACAGTGGTCTCGGATAGGATATATTAACCCAAGTTCTGCCTACACTAACTATA CAACCCCCCTACTTTCGGGGGATCAGAAGTTTCAAGGCCGGGTAACACTTACAGCCGATAA AGGTACAAAGGTAGAAATTAAGAGTACCTCAACAGCATACATGGAACTTAGTTCTTTGCGG GGGCGGCTCCGAGGGCAAGAAGCGAGGATACCGCTGTGTATTACTGCGCTTCACCTCAGG GCAGCGGCAGCGGCAGCGAGTTCACTACGACTACGCTGGATTTCCCTATTGGGGTCAGGGT AGCAAGAGCACCGGCGGCAGACACTGGTTACAGTTTCCTCTGAGCCCAAATCTAGCGACAA CCAAGTGCAACTCGTGCAAAGAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCA CGGGGCTGAAGTGAAGAAGCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGG CTGGATCAAGCGTGAAGGTCAACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTG GTTGCAAAGCCTCTGGATATAGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTC CCTTCACTCGATCAACCATGCAAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAG CTGGGTCAAGCAGGCCCCAGGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGT GCAAGGGCTCGAATGGATAGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA GATATATTAACCCAAGTTCTGCATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCT CTACACTAACTATAATCAGAACCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGG GTTTCAAGGCCGGGTAACACTCAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCC TACAGCCGATAAGAGTACCTCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCC AACAGCATACATGGAACTTAGTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTG TTCTTTGCGGAGCGAGGATACGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCAC CGCTGTGTATTACTGCGCTTCAGCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGA CCTCAGGTTCACTACGACTACGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGA GCTGGATTTCCCTATTGGGGTACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC CAGGGTACACTGGTTACAGTTCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT TCCTCTG (182) (183) CD3B2030-GAAATTGTCTTGACCCAGTCTCCAGCAACTCTTAGTGCATC GAAATTGTCTTGACCCAGTCTC LH-scFvACCAGGTGAGCGTGTTACCCTCTCATGTAGCGCCAGCTCAT CAGCAACTCTTAGTGCATCACCCTGTTAGTTATATGAATTGGTATCAACAGAAACCAGGGCA AGGTGAGCGTGTTACCCTCTCAGCTCCCAGAAGATGGATATATGATTCTTCAAAACTCGCA ATGTAGCGCCAGCTCATCTGTTAGTGGTGTCCCAGCCCGCTTCTCAGGCTCTGGTTCCGGTC AGTTATATGAATTGGTATCAAGCGATTATACTCTCACCATCAGTAGTTTGGAACCCGAAGAT CAGAAACCAGGGCAAGCTCCCTTCGCCGTCTATTATTGCCAGCAATGGAGCAGGAATCCCCC AGAAGATGGATATATGATTCTCACATTCGGCGGCGGTACAAAGGTTGAGATTAAGGGCGG TCAAAACTCGCAAGTGGTGTCCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCA CCAGCCCGCTTCTCAGGCTCTGAGAGCACCGGCGGCAGCCAAGTTCAGTTGGTTCAATCCGG GTTCCGGTCGCGATTATACTCTCGCAGAGGTTAAAAAACCCGGATCAAGCGTTAAGGTTAGT CACCATCAGTAGTTTGGAACCTGTAAAGCCTCTGGCTACACTTTCACACGCTCAACAATGCA CGAAGATTTCGCCGTCTATTATTTGGGTTAAGCAGGCCCCTGGGCAGGGACTGGAGTGGAT TGCCAGCAATGGAGCAGGAATCGGTTACATAAACCCATCCAGCGCCTATACAAACTATAACC CCCCCCACATTCGGCGGCGGTAGAAGTTCCAAGGGCGGGTTACATTGACCGCTGACAAGTC ACAAAGGTTGAGATTAAGGGCCACTAGCACAGCATATATGGAGCTGTCAAGTCTGAGATCC GGCTCCGAGGGCAAGAGCAGGAAGACACTGCCGTATATTATTGCGCTAGTCCACAAGTGC CGGCAGCGGCAGCGAGAGCAACTATGACTATAACGGTTTTCCCTATTGGGGACAAGGAAC AGAGCACCGGCGGCAGCCAACCTGGTGACCGTTAGCTCCGAGCCCAAATCTAGCGACAAA GTTCAGTTGGTTCAATCCGGCACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG GCAGAGGTTAAAAAACCCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC ATCAAGCGTTAAGGTTAGTTGACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT TAAAGCCTCTGGCTACACTTTCGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA ACACGCTCAACAATGCATTGGCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC GTTAAGCAGGCCCCTGGGCAGAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT GGACTGGAGTGGATCGGTTACGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT ATAAACCCATCCAGCGCCTATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC ACAAACTATAACCAGAAGTTCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA CAAGGGCGGGTTACATTGACCGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG GCTGACAAGTCCACTAGCACAGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GCATATATGGAGCTGTCAAGTGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG CTGAGATCCGAAGACACTGCCAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC GTATATTATTGCGCTAGTCCACCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC AAGTGCACTATGACTATAACGAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC GTTTTCCCTATTGGGGACAAGGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GAACCCTGGTGACCGTTAGCTCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT CCG (184) (185) CD3B2089-CAAGTGCAGTTGGTCCAGTCAGGCGCAGAAGTGAAGAAG CAAGTGCAGTTGGTCCAGTCA N106S-CCCGGCTCAAGCGTCAAGGTATCATGTAAGGCTTCTGGAT GGCGCAGAAGTGAAGAAGCC HL-scFvATACTTTCACCCGAAGCACAATGCACTGGGTTCGCCAGGC CGGCTCAAGCGTCAAGGTATCCCCTGGACAGGGTCTTGAGTGGATGGGGTATATCAACCCA ATGTAAGGCTTCTGGATATACTCCTCAGCATATACTAACTATGCTCAGAAGTTTCAGGGGCG TTTCACCCGAAGCACAATGCATGTCACTTTGACCGCCGATAAGTCCACAAGCACCGCTTATA CTGGGTTCGCCAGGCCCCTGGTGGAACTGTCTTCATTGCGCTCTGAAGACACTGCAGTGTAC ACAGGGTCTTGAGTGGATGGTATTGCGCCAGCCCACAGGTCCACTACGACTATTCTGGATT GGTATATCAACCCATCCTCAGCTCCATACTGGGGGCAGGGGACCTTGGTGACTGTAAGCTCT ATATACTAACTATGCTCAGAAGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGA GTTTCAGGGGCGTGTCACTTTGAGCAAGAGCACCGGCGGCAGCGAGATTGTACTGACACA GACCGCCGATAAGTCCACAAGGTCCCCAGCAACCTTGTCCGCTTCTCCCGGCGAAAGGGTC CACCGCTTATATGGAACTGTCTACTCTCTCCTGCTCCGCTAGTTCTTCAGTGTCATATATGAAT TCATTGCGCTCTGAAGACACTTGGTACCAACAAAAGCCAGGTCAGGCTCCAAGAAGATGG GCAGTGTACTATTGCGCCAGCATTTACGATTCCTCCAAGTTGGCTTCTGGTGTCCCTGCACG CCACAGGTCCACTACGACTATTATTTAGCGGGTCAGGGTCAGGGCGCGATTACACACTCACA CTGGATTTCCATACTGGGGGCATTAGTAGTCTCGAACCCGAGGACTTTGCCGTATATTACTG AGGGGACCTTGGTGACTGTAATCAGCAATGGAGTCGGAATCCCCCAACTTTCGGCGGGGGA GCTCTGGCGGCTCCGAGGGCAACAAAAGTAGAAATAAAAGAGCCCAAATCTAGCGACAAA AGAGCAGCGGCAGCGGCAGCACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG GAGAGCAAGAGCACCGGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC CAGCGAGATTGTACTGACACAACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT GTCCCCAGCAACCTTGTCCGCTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA TCTCCCGGCGAAAGGGTCACTCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC CTCTCCTGCTCCGCTAGTTCTTAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT CAGTGTCATATATGAATTGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT ACCAACAAAAGCCAGGTCAGGGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC CTCCAAGAAGATGGATTTACGCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA ATTCCTCCAAGTTGGCTTCTGGGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG TGTCCCTGCACGATTTAGCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GTCAGGGTCAGGGCGCGATTAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG CACACTCACAATTAGTAGTCTCAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC GAACCCGAGGACTTTGCCGTACTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC TATTACTGTCAGCAATGGAGTAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC CGGAATCCCCCAACTTTCGGCGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GGGGGAACAAAAGTAGAAATCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AAAAG (186) (187) CD3B2089-CAAGTGCAGTTGGTTCAATCCGGCGCTGAAGTGAAGAAAC CAAGTGCAGTTGGTTCAATCC N106Q-CTGGGTCATCTGTCAAAGTATCCTGTAAAGCCTCTGGGTAC GGCGCTGAAGTGAAGAAACCT HL-scFvACTTTTACACGTAGCACCATGCACTGGGTCCGTCAAGCCCC GGGTCATCTGTCAAAGTATCCTGGGCAAGGCCTTGAGTGGATGGGTTATATAAACCCATCC TGTAAAGCCTCTGGGTACACTTCCGCATACACAAATTACGCTCAAAAATTTCAAGGGCGAG TTTACACGTAGCACCATGCACTTCACTCTCACTGCCGATAAATCCACTTCAACTGCCTATATG GGGTCCGTCAAGCCCCTGGGCGAGCTTAGTTCATTGCGATCAGAAGATACTGCAGTCTATTA AAGGCCTTGAGTGGATGGGTTTTGTGCATCACCTCAGGTCCATTACGACTACCAAGGGTTCC ATATAAACCCATCCTCCGCATACCTACTGGGGACAGGGGACTTTGGTAACTGTGTCTTCTGG CACAAATTACGCTCAAAAATTTCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGA CAAGGGCGAGTCACTCTCACTGCAAGAGCACCGGCGGCAGCGAGATCGTACTGACCCAAA GCCGATAAATCCACTTCAACTGTCCCGCCACTCTCTCTGCTAGCCCAGGCGAGAGAGTTAC GCCTATATGGAGCTTAGTTCATCTTGTCTTGCTCTGCTAGTTCAAGTGTCAGTTATATGAACT TGCGATCAGAAGATACTGCAGGGTATCAGCAGAAGCCAGGACAGGCACCTCGAAGATGGA TCTATTATTGTGCATCACCTCATATATGACTCCTCCAAACTCGCATCAGGCGTACCAGCACGC GGTCCATTACGACTACCAAGGTTTTCTGGGAGCGGTAGTGGTAGGGATTATACACTCACCA GTTCCCCTACTGGGGACAGGGTCTCTAGTTTGGAACCAGAAGATTTCGCTGTGTACTATTGC GACTTTGGTAACTGTGTCTTCTCAGCAGTGGAGCCGCAACCCTCCTACCTTCGGCGGTGGGA GGCGGCTCCGAGGGCAAGAGCAAAGGTAGAAATAAAAGAGCCCAAATCTAGCGACAAAA CAGCGGCAGCGGCAGCGAGACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGG GCAAGAGCACCGGCGGCAGCGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACA GAGATCGTACTGACCCAAAGTCCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT CCCGCCACTCTCTCTGCTAGCCGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA CAGGCGAGAGAGTTACCTTGTCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC CTTGCTCTGCTAGTTCAAGTGTAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT CAGTTATATGAACTGGTATCAGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT GCAGAAGCCAGGACAGGCACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC CTCGAAGATGGATATATGACTCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA CCTCCAAACTCGCATCAGGCGGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG TACCAGCACGCTTTTCTGGGAGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GCGGTAGTGGTAGGGATTATAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG CACTCACCATCTCTAGTTTGGAAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC ACCAGAAGATTTCGCTGTGTACTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC CTATTGCCAGCAGTGGAGCCGAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC CAACCCTCCTACCTTCGGCGGTGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GGGACAAAGGTAGAAATAAACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AG (188) (189) CD3B2089-CAAGTTCAGCTTGTGCAGAGCGGGGCAGAGGTGAAGAAA CAAGTTCAGCTTGTGCAGAGC N106G-CCCGGATCAAGCGTCAAAGTTTCTTGTAAAGCTAGTGGAT GGGGCAGAGGTGAAGAAACC HL-scFvATACTTTCACACGCTCAACTATGCACTGGGTGAGACAAGCT CGGATCAAGCGTCAAAGTTTCCCTGGTCAGGGCCTGGAGTGGATGGGGTACATAAATCCCT TTGTAAAGCTAGTGGATATACCCAGTGCATATACTAACTATGCTCAAAAGTTCCAAGGCCGC TTTCACACGCTCAACTATGCACGTAACTCTCACTGCCGATAAGTCCACCAGCACTGCCTACAT TGGGTGAGACAAGCTCCTGGTGGAACTGTCTAGTTTGCGATCCGAGGACACCGCCGTGTAC CAGGGCCTGGAGTGGATGGGTACTGTGCTTCACCTCAAGTACATTATGACTACGGGGGATT GTACATAAATCCCTCCAGTGCTCCCTACTGGGGCCAAGGTACTTTGGTCACAGTCTCAAGC ATATACTAACTATGCTCAAAAGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGA GTTCCAAGGCCGCGTAACTCTGAGCAAGAGCACCGGCGGCAGCGAGATCGTATTGACACA CACTGCCGATAAGTCCACCAGATCACCCGCCACATTGTCAGCTAGCCCCGGTGAGCGCGTC CACTGCCTACATGGAACTGTCTACACTTTCTTGTAGTGCATCAAGTAGCGTTTCTTACATGAA AGTTTGCGATCCGAGGACACCTTGGTATCAGCAGAAACCAGGACAAGCACCACGGCGATG GCCGTGTACTACTGTGCTTCACGATATACGATTCTAGCAAACTCGCCAGTGGCGTCCCCGCTC CTCAAGTACATTATGACTACGGATTCTCCGGGTCTGGCAGTGGTAGAGATTATACACTCAC GGGGATTTCCCTACTGGGGCCTATCAGTTCTCTGGAACCAGAAGACTTCGCAGTCTATTACT AAGGTACTTTGGTCACAGTCTGTCAACAATGGTCACGGAATCCCCCCACATTCGGTGGTGG CAAGCGGCGGCTCCGAGGGCCACCAAGGTTGAAATTAAGGAGCCCAAATCTAGCGACAAA AAGAGCAGCGGCAGCGGCAGACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG CGAGAGCAAGAGCACCGGCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC GCAGCGAGATCGTATTGACACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT AATCACCCGCCACATTGTCAGGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA CTAGCCCCGGTGAGCGCGTCACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC CACTTTCTTGTAGTGCATCAAGAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT TAGCGTTTCTTACATGAATTGGGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT TATCAGCAGAAACCAGGACAAGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC GCACCACGGCGATGGATATACCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA GATTCTAGCAAACTCGCCAGTGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG GGCGTCCCCGCTCGATTCTCCGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GGGTCTGGCAGTGGTAGAGAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG TTATACACTCACTATCAGTTCTAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC CTGGAACCAGAAGACTTCGCACTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC GTCTATTACTGTCAACAATGGTAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC CACGGAATCCCCCCACATTCGGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GTGGTGGCACCAAGGTTGAAACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT TTAAGG (190) (191) CD3B2089-CAAGTGCAATTGGTCCAAAGTGGAGCTGAAGTAAAAAAA CAAGTGCAATTGGTCCAAAGT N106A-CCCGGCTCCTCTGTGAAGGTCAGTTGCAAAGCCTCAGGGT GGAGCTGAAGTAAAAAAACCC HL-scFvACACCTTTACTAGGTCAACAATGCACTGGGTGCGACAAGC GGCTCCTCTGTGAAGGTCAGTTCCCGGTCAGGGTTTGGAGTGGATGGGATACATAAACCCC TGCAAAGCCTCAGGGTACACCTCATCAGCCTACACAAATTATGCACAAAAATTTCAGGGTCG TTTACTAGGTCAACAATGCACTGGTTACACTCACCGCCGACAAATCCACTTCCACTGCTTATA GGGTGCGACAAGCTCCCGGTCTGGAACTTTCCTCTCTCCGCAGTGAGGACACAGCAGTGTA AGGGTTTGGAGTGGATGGGACTATTGTGCCTCCCCTCAAGTGCATTATGACTACGCTGGTT TACATAAACCCCTCATCAGCCTTCCCTTACTGGGGACAAGGTACTCTGGTTACAGTTTCTTCC ACACAAATTATGCACAAAAATGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGA TTCAGGGTCGGGTTACACTCAGAGCAAGAGCACCGGCGGCAGCGAAATAGTGCTGACCCA CCGCCGACAAATCCACTTCCACGAGCCCCGCTACCCTTTCTGCAAGTCCTGGGGAACGTGTT TGCTTATATGGAACTTTCCTCTACATTGTCTTGTAGCGCTTCTTCATCAGTCTCCTATATGAAT CTCCGCAGTGAGGACACAGCATGGTATCAACAAAAACCAGGACAAGCTCCTCGGCGGTGGA GTGTACTATTGTGCCTCCCCTCTCTACGACAGTTCCAAACTTGCCTCTGGTGTGCCTGCTCGG AAGTGCATTATGACTACGCTGTTTAGTGGGTCTGGAAGTGGACGAGATTATACTCTGACCA GTTTCCCTTACTGGGGACAAGTCAGTTCCTTGGAACCCGAGGATTTTGCTGTTTATTACTGC GTACTCTGGTTACAGTTTCTTCCAACAATGGAGTAGAAACCCTCCAACCTTTGGAGGTGGAA CGGCGGCTCCGAGGGCAAGACTAAGGTCGAGATAAAGGAGCCCAAATCTAGCGACAAAA GCAGCGGCAGCGGCAGCGAGCTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGG AGCAAGAGCACCGGCGGCAGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACA CGAAATAGTGCTGACCCAGAGCCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT CCCCGCTACCCTTTCTGCAAGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA CCTGGGGAACGTGTTACATTGCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC TCTTGTAGCGCTTCTTCATCAGAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT TCTCCTATATGAATTGGTATCAGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT ACAAAAACCAGGACAAGCTCCGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC TCGGCGGTGGATCTACGACAGCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA TTCCAAACTTGCCTCTGGTGTGGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG CCTGCTCGGTTTAGTGGGTCTGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GGAAGTGGACGAGATTATACTGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG CTGACCATCAGTTCCTTGGAACAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC CCGAGGATTTTGCTGTTTATTACTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC CTGCCAACAATGGAGTAGAAAAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC CCCTCCAACCTTTGGAGGTGGGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA AACTAAGGTCGAGATAAAGGCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT (193) (192) CD3B2089-CAGGTACAACTCGTGCAAAGTGGTGCTGAAGTGAAGAAA CAGGTACAACTCGTGCAAAGT HL-scFvCCTGGATCAAGCGTCAAGGTATCCTGTAAAGCATCAGGAT GGTGCTGAAGTGAAGAAACCTACACCTTCACACGCAGTACTATGCATTGGGTGCGTCAAGC GGATCAAGCGTCAAGGTATCCCCCCGGACAGGGCCTGGAATGGATGGGCTACATAAACCCT TGTAAAGCATCAGGATACACCTCTTCCGCCTACACCAATTATGCCCAAAAGTTCCAGGGAAG TTCACACGCAGTACTATGCATTGGTGACTCTGACTGCTGATAAAAGTACTAGCACCGCATAC GGGTGCGTCAAGCCCCCGGACATGGAACTGTCTTCACTGAGAAGCGAGGACACCGCCGTCT AGGGCCTGGAATGGATGGGCATTATTGTGCATCCCCCCAAGTCCACTATGATTACAACGGA TACATAAACCCTTCTTCCGCCTTTTCCTTACTGGGGCCAGGGAACCTTGGTCACCGTGTCTTC ACACCAATTATGCCCAAAAGTCGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCG TCCAGGGAAGGGTGACTCTGAAGAGCAAGAGCACCGGCGGCAGCGAAATCGTTCTCACAC CTGCTGATAAAAGTACTAGCAAGAGCCCTGCAACATTGTCAGCTTCACCCGGTGAACGAGT CCGCATACATGGAACTGTCTTCAACATTGTCCTGTTCTGCCTCAAGTAGTGTGAGCTATATGA ACTGAGAAGCGAGGACACCGATTGGTATCAACAAAAACCAGGGCAGGCCCCTAGAAGGT CCGTCTATTATTGTGCATCCCCGGATCTATGATTCAAGCAAACTGGCATCCGGCGTCCCTGC CCAAGTCCACTATGATTACAACCCGCTTTAGTGGAAGCGGTTCAGGAAGGGACTATACTCTT GGATTTCCTTACTGGGGCCAGACTATCTCCAGCCTTGAACCTGAAGATTTTGCAGTCTACTA GGAACCTTGGTCACCGTGTCTCTGCCAACAATGGTCTAGGAATCCCCCCACTTTTGGTGGA TCCGGCGGCTCCGAGGGCAAGGGACCAAAGTTGAGATCAAAGAGCCCAAATCTAGCGAC GAGCAGCGGCAGCGGCAGCGAAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAG AGAGCAAGAGCACCGGCGGCCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAG AGCGAAATCGTTCTCACACAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGT AGCCCTGCAACATTGTCAGCTTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTT CACCCGGTGAACGAGTAACATCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAA TGTCCTGTTCTGCCTCAAGTAGGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCG TGTGAGCTATATGAATTGGTATGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTG TCAACAAAAACCAGGGCAGGCAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCC CCCTAGAAGGTGGATCTATGACTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAG TTCAAGCAAACTGGCATCCGGGGCAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATC CGTCCCTGCCCGCTTTAGTGGCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTG AAGCGGTTCAGGAAGGGACTCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAG ATACTCTTACTATCTCCAGCCTTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACC TGAACCTGAAGATTTTGCAGTACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGT CTACTACTGCCAACAATGGTCTGAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGG AGGAATCCCCCCACTTTTGGTGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACA GGAGGGACCAAAGTTGAGATACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT CAAAG (194) (195) CD3B2051-CAAGTGCAACTCGTACAATCTGGCGCTGAGGTTAAGAAAC CAAGTGCAACTCGTACAATCT N106S-CTGGTAGCTCTGTTAAAGTGTCTTGTAAAGCATCCGGGTAT GGCGCTGAGGTTAAGAAACCT HL-scFvACTTTTACCCGGTCAACTATGCACTGGGTAAAACAAGCTCC GGTAGCTCTGTTAAAGTGTCTTTGGACAAGGTTTGGAGTGGATGGGTTATATAAATCCCTCC GTAAAGCATCCGGGTATACTTTCAGCATACACTAACTACAACCAGAAGTTCCAGGGGCGCG TTACCCGGTCAACTATGCACTGTTACCCTGACTGCCGATAAGAGTACTTCAACTGCTTATATG GGTAAAACAAGCTCCTGGACAGAGCTGTCATCCCTGCGTAGCGAGGACACAGCAGTATACT AGGTTTGGAGTGGATGGGTTAACTGCGCCAGTCCACAGGTACACTACGATTACAGTGGCTTT TATAAATCCCTCCTCAGCATACCCATACTGGGGGCAGGGCACTCTGGTAACAGTATCTAGTG ACTAACTACAACCAGAAGTTCGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAG CAGGGGCGCGTTACCCTGACTAGCAAGAGCACCGGCGGCAGCGAGATTGTACTCACCCAG GCCGATAAGAGTACTTCAACTTCTCCAGCTACCCTTAGTGCTTCACCTGGTGAGCGCGTGAC GCTTATATGGAGCTGTCATCCCATTGTCCTGCTCCGCAAGCTCCAGTGTTTCATATATGAATT TGCGTAGCGAGGACACAGCAGGTACCAACAAAAGCCTGGGCAAGCACCACGCCGGCTGA GTATACTACTGCGCCAGTCCATCTACGACAGCTCCAAGCTCGCAAGCGGTGTACCTGCTCG CAGGTACACTACGATTACAGTCTTTTCCGGCAGCGGGTCAGGTCGAGATTATACTCTGACC GGCTTTCCATACTGGGGGCAGATTTCATCACTCGAACCCGAAGACTTTGCAGTGTATTACTG GGCACTCTGGTAACAGTATCTTCAACAGTGGAGTAGGAATCCACCAACATTTGGGGGTGGC AGTGGCGGCTCCGAGGGCAAACCAAGGTTGAGATAAAGGAGCCCAAATCTAGCGACAAA GAGCAGCGGCAGCGGCAGCGACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG AGAGCAAGAGCACCGGCGGCGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC AGCGAGATTGTACTCACCCAGACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT TCTCCAGCTACCCTTAGTGCTTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA CACCTGGTGAGCGCGTGACATCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC TGTCCTGCTCCGCAAGCTCCAAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT GTGTTTCATATATGAATTGGTAGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT CCAACAAAAGCCTGGGCAAGCGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC ACCACGCCGGCTGATCTACGACAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA CAGCTCCAAGCTCGCAAGCGGGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG TGTACCTGCTCGCTTTTCCGGCGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG AGCGGGTCAGGTCGAGATTATGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG ACTCTGACCATTTCATCACTCGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC AACCCGAAGACTTTGCAGTGTCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC ATTACTGTCAACAGTGGAGTAAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC GGAATCCACCAACATTTGGGGGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GTGGCACCAAGGTTGAGATAACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AGG (196) (197) CD3B2051-CAGGTACAGTTGGTCCAAAGTGGCGCAGAGGTAAAGAAA CAGGTACAGTTGGTCCAAAGT N106Q-CCAGGTTCTTCAGTCAAGGTAAGTTGCAAGGCATCTGGAT GGCGCAGAGGTAAAGAAACC HL-scFvATACATTTACCCGCAGTACTATGCATTGGGTCAAACAGGCT AGGTTCTTCAGTCAAGGTAAGCCAGGACAGGGGCTTGAATGGATGGGTTACATCAACCCAT TTGCAAGGCATCTGGATATACCTAGTGCCTATACAAACTATAATCAGAAATTTCAGGGCAG ATTTACCCGCAGTACTATGCATAGTGACTCTGACAGCCGACAAATCAACCTCTACAGCATAT TGGGTCAAACAGGCTCCAGGAATGGAGTTGTCCTCTCTCCGTAGTGAAGATACTGCCGTCTA CAGGGGCTTGAATGGATGGGCTATTGTGCAAGCCCCCAAGTCCACTATGATTATCAGGGTT TTACATCAACCCATCTAGTGCCTCCCTTACTGGGGGCAGGGTACTTTGGTTACCGTTTCATCC TATACAAACTATAATCAGAAAGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGA TTTCAGGGCAGAGTGACTCTGGAGCAAGAGCACCGGCGGCAGCGAGATCGTGTTGACTCA ACAGCCGACAAATCAACCTCTAAGTCCTGCAACCCTGTCTGCTAGTCCAGGGGAGAGGGTT ACAGCATATATGGAGTTGTCCACTCTCAGTTGTTCTGCAAGCAGTAGCGTATCCTACATGAA TCTCTCCGTAGTGAAGATACTCTGGTATCAACAAAAGCCTGGTCAGGCACCACGGCGGTTG GCCGTCTACTATTGTGCAAGCATATATGACTCCTCCAAGTTGGCCTCTGGGGTGCCCGCAA CCCCAAGTCCACTATGATTATCGATTCTCCGGGTCCGGCTCTGGCCGCGATTACACACTGAC AGGGTTTCCCTTACTGGGGGCTATAAGCAGTCTGGAACCAGAGGATTTTGCCGTTTACTACT AGGGTACTTTGGTTACCGTTTCGCCAACAATGGAGCCGAAACCCCCCAACCTTTGGAGGTGG ATCCGGCGGCTCCGAGGGCAACACTAAGGTAGAGATAAAGGAGCCCAAATCTAGCGACAA GAGCAGCGGCAGCGGCAGCGAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCA AGAGCAAGAGCACCGGCGGCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGG AGCGAGATCGTGTTGACTCAAACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTG AGTCCTGCAACCCTGTCTGCTAGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTC GTCCAGGGGAGAGGGTTACTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAG TCAGTTGTTCTGCAAGCAGTAACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGT GCGTATCCTACATGAACTGGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA ATCAACAAAAGCCTGGTCAGGATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCT CACCACGGCGGTTGATATATGCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGG ACTCCTCCAAGTTGGCCTCTGGCAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCC GGTGCCCGCAAGATTCTCCGGGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCC GTCCGGCTCTGGCCGCGATTATGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTG CACACTGACTATAAGCAGTCTGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCAC GGAACCAGAGGATTTTGCCGTGCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGA TTACTACTGCCAACAATGGAGGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGA CCGAAACCCCCCAACCTTTGGACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC AGGTGGCACTAAGGTAGAGACACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT TAAAGG (198) (199) CD3B2051-CAAGTTCAACTGGTCCAAAGCGGTGCTGAAGTTAAAAAGC CAAGTTCAACTGGTCCAAAGC N106G-CAGGAAGCAGTGTTAAAGTCTCATGTAAGGCCAGCGGTTA GGTGCTGAAGTTAAAAAGCCA HL-scFvCACTTTTACTAGGAGTACCATGCACTGGGTGAAGCAGGCC GGAAGCAGTGTTAAAGTCTCACCCGGTCAGGGTCTTGAGTGGATGGGATATATAAACCCAT TGTAAGGCCAGCGGTTACACTCATCCGCCTACACTAATTACAACCAAAAGTTTCAGGGTCGC TTTACTAGGAGTACCATGCACTGTGACTTTGACCGCCGACAAATCTACCAGCACAGCCTACAT GGGTGAAGCAGGCCCCCGGTGGAACTCAGTTCTCTCCGATCCGAAGATACCGCTGTATATT CAGGGTCTTGAGTGGATGGGACTGTGCTTCCCCACAAGTACACTATGATTACGGGGGCTTC ATATATAAACCCATCATCCGCCCCATACTGGGGCCAGGGAACTCTCGTCACAGTATCATCCG TACACTAATTACAACCAAAAGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAG TTTCAGGGTCGCGTGACTTTGAGCAAGAGCACCGGCGGCAGCGAAATTGTTCTTACACAAA ACCGCCGACAAATCTACCAGCGTCCTGCTACACTGTCAGCCAGCCCCGGTGAGCGAGTCAC ACAGCCTACATGGAACTCAGTATTGTCATGCTCTGCTTCCAGTAGTGTGAGCTACATGAACT TCTCTCCGATCCGAAGATACCGGTACCAACAGAAACCTGGTCAGGCTCCAAGGCGCTTGAT GCTGTATATTACTGTGCTTCCCATACGACAGCAGCAAACTGGCAAGTGGTGTACCTGCTCGG CACAAGTACACTATGATTACGTTTTCTGGATCAGGCTCAGGTAGAGACTATACTCTCACCAT GGGGCTTCCCATACTGGGGCCTTCCTCTCTGGAACCTGAGGACTTTGCTGTTTATTATTGCCA AGGGAACTCTCGTCACAGTATGCAGTGGAGTCGCAACCCTCCCACCTTCGGTGGAGGGACA CATCCGGCGGCTCCGAGGGCAAAAGTAGAAATAAAGGAGCCCAAATCTAGCGACAAAACTC AGAGCAGCGGCAGCGGCAGCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGG GAGAGCAAGAGCACCGGCGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACC CAGCGAAATTGTTCTTACACACTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGG AAGTCCTGCTACACTGTCAGCTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTG CAGCCCCGGTGAGCGAGTCACGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAA ATTGTCATGCTCTGCTTCCAGTGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGT AGTGTGAGCTACATGAACTGGCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGC TACCAACAGAAACCTGGTCAGAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAG GCTCCAAGGCGCTTGATATACCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCC GACAGCAGCAAACTGGCAAGTCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGGGAG GGTGTACCTGCTCGGTTTTCTGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCA GATCAGGCTCAGGTAGAGACTAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGA ATACTCTCACCATTTCCTCTCTGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTC GGAACCTGAGGACTTTGCTGTCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGCAA TTATTATTGCCAGCAGTGGAGGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGT TCGCAACCCTCCCACCTTCGGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACT GGAGGGACAAAAGTAGAAATACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AAAGG (200) (201) CD3B2051-CAAGTTCAGTTGGTTCAATCCGGCGCTGAGGTCAAAAAAC CAAGTTCAGTTGGTTCAATCC N106A-CTGGATCATCTGTGAAAGTCTCATGTAAGGCATCTGGTTAT GGCGCTGAGGTCAAAAAACCT HL-scFvACCTTCACTCGGAGTACCATGCATTGGGTTAAGCAGGCCC GGATCATCTGTGAAAGTCTCACCGGTCAGGGGTTGGAGTGGATGGGTTACATCAACCCTTC TGTAAGGCATCTGGTTATACCTCTCAGCCTACACAAATTATAATCAGAAATTTCAGGGGCGC TCACTCGGAGTACCATGCATTGTTACTCTCACCGCTGACAAGTCCACCTCCACAGCCTATAT GGGTTAAGCAGGCCCCCGGTCGGAGCTGTCAAGCCTGCGGAGTGAGGATACAGCCGTATA AGGGGTTGGAGTGGATGGGTTTACTGTGCCAGTCCTCAGGTTCATTATGATTACGCTGGCT TACATCAACCCTTCCTCAGCCTTCCCATATTGGGGTCAGGGGACTCTCGTCACTGTGTCCAG ACACAAATTATAATCAGAAATCGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCG TTCAGGGGCGCGTTACTCTCAAGAGCAAGAGCACCGGCGGCAGCGAAATTGTATTGACTC CCGCTGACAAGTCCACCTCCAAGTCCCCAGCTACATTGAGCGCAAGTCCTGGCGAGAGAGT CAGCCTATATGGAGCTGTCAAAACCCTGTCTTGTTCTGCCAGTAGTAGTGTAAGCTACATGA GCCTGCGGAGTGAGGATACAACTGGTATCAGCAGAAACCCGGACAGGCCCCACGCCGACT GCCGTATATTACTGTGCCAGTCTATCTATGATTCAAGTAAGCTCGCTAGTGGGGTTCCAGCCA CTCAGGTTCATTATGATTACGCGATTTAGTGGTTCTGGCTCTGGACGCGATTACACTCTGACC TGGCTTCCCATATTGGGGTCAATTTCTTCTCTGGAGCCTGAGGACTTCGCAGTATATTACTG GGGGACTCTCGTCACTGTGTCCCAACAATGGTCACGCAATCCACCAACATTCGGTGGAGGG CAGCGGCGGCTCCGAGGGCAACAAAAGTGGAAATCAAAGAGCCCAAATCTAGCGACAAA AGAGCAGCGGCAGCGGCAGCACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG GAGAGCAAGAGCACCGGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC CAGCGAAATTGTATTGACTCAACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT GTCCCCAGCTACATTGAGCGCGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA AAGTCCTGGCGAGAGAGTAACCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC CCTGTCTTGTTCTGCCAGTAGTAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT AGTGTAAGCTACATGAACTGGGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT TATCAGCAGAAACCCGGACAGGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC GCCCCACGCCGACTTATCTATGCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA ATTCAAGTAAGCTCGCTAGTGGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG GGGTTCCAGCCAGATTTAGTGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GTTCTGGCTCTGGACGCGATTGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG ACACTCTGACCATTTCTTCTCTAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC GGAGCCTGAGGACTTCGCAGTCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC ATATTACTGCCAACAATGGTCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC ACGCAATCCACCAACATTCGGGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA TGGAGGGACAAAAGTGGAAACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT TCAAAG (202) (203) CD3B2051-CAAGTGCAGTTGGTGCAGAGTGGGGCTGAGGTTAAAAAG CAAGTGCAGTTGGTGCAGAGT HL-scFvCCTGGTTCCAGTGTGAAAGTCAGTTGTAAAGCCTCCGGGT GGGGCTGAGGTTAAAAAGCCTACACTTTTACTAGGTCAACAATGCACTGGGTCAAGCAAGC GGTTCCAGTGTGAAAGTCAGTCCCCGGCCAAGGCTTGGAATGGATGGGGTACATAAATCCA TGTAAAGCCTCCGGGTACACTAGCAGTGCCTACACCAACTATAACCAAAAATTTCAAGGTA TTTACTAGGTCAACAATGCACTGAGTAACATTGACTGCTGACAAGTCCACATCAACTGCTTAT GGGTCAAGCAAGCCCCCGGCCATGGAGCTGTCCTCTCTTCGGTCTGAAGATACCGCCGTATA AAGGCTTGGAATGGATGGGGCTATTGCGCCTCCCCCCAAGTCCACTACGACTATAACGGAT TACATAAATCCAAGCAGTGCCTTCCCTACTGGGGACAAGGAACCCTGGTAACAGTTTCTTCA TACACCAACTATAACCAAAAAGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGA TTTCAAGGTAGAGTAACATTGGAGCAAGAGCACCGGCGGCAGCGAGATAGTTCTTACACA ACTGCTGACAAGTCCACATCAGAGCCCTGCAACCTTGAGTGCAAGTCCAGGGGAACGGGT ACTGCTTATATGGAGCTGTCCTGACTCTGAGTTGTAGTGCTTCTAGTTCCGTAAGTTATATGA CTCTTCGGTCTGAAGATACCGACTGGTACCAACAGAAGCCAGGTCAAGCACCAAGACGCCT CCGTATACTATTGCGCCTCCCCTATCTACGACTCATCTAAACTTGCTAGTGGAGTGCCAGCCA CCAAGTCCACTACGACTATAAGATTTTCCGGTTCAGGAAGTGGGAGGGACTACACACTTAC CGGATTTCCCTACTGGGGACACATCTCATCCCTTGAGCCCGAAGATTTCGCCGTATATTACT AGGAACCCTGGTAACAGTTTCGTCAACAATGGTCAAGAAATCCTCCTACATTTGGTGGTGG TTCAGGCGGCTCCGAGGGCAATACAAAAGTAGAGATCAAGGAGCCCAAATCTAGCGACAAA GAGCAGCGGCAGCGGCAGCGACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG AGAGCAAGAGCACCGGCGGCGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC AGCGAGATAGTTCTTACACAGACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT AGCCCTGCAACCTTGAGTGCAGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA AGTCCAGGGGAACGGGTGACCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC TCTGAGTTGTAGTGCTTCTAGTAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT TCCGTAAGTTATATGAACTGGGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT TACCAACAGAAGCCAGGTCAAGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC GCACCAAGACGCCTTATCTACCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA GACTCATCTAAACTTGCTAGTGGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG GAGTGCCAGCCAGATTTTCCGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GTTCAGGAAGTGGGAGGGACGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG TACACACTTACCATCTCATCCCAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC TTGAGCCCGAAGATTTCGCCGCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC TATATTACTGTCAACAATGGTCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC AAGAAATCCTCCTACATTTGGTGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GGTGGTACAAAAGTAGAGATCCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AAGG (204) (205) CD3B2030-CAAGTACAGCTCGTTCAGTCCGGTGCAGAAGTCAAGAAAC CAAGTACAGCTCGTTCAGTCC N106S-CAGGAAGTAGCGTAAAAGTGTCATGTAAAGCAAGTGGTT GGTGCAGAAGTCAAGAAACC HL-scFvATACCTTTACACGCTCAACTATGCATTGGGTTAAGCAGGCT AGGAAGTAGCGTAAAAGTGTCCCAGGACAAGGGCTTGAGTGGATAGGATACATCAATCCAT ATGTAAAGCAAGTGGTTATACCTAGCGCCTACACAAATTATAACCAGAAGTTCCAGGGGAG CTTTACACGCTCAACTATGCATAGTTACCCTCACTGCCGATAAGTCCACATCAACCGCCTATA TGGGTTAAGCAGGCTCCAGGATGGAATTGAGTTCCCTTCGTAGTGAGGACACTGCCGTCTA CAAGGGCTTGAGTGGATAGGCTACTGTGCCTCCCCTCAGGTTCATTATGATTACTCAGGTTT ATACATCAATCCATCTAGCGCCTCCATACTGGGGCCAGGGCACCCTCGTAACAGTAAGCAGC TACACAAATTATAACCAGAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGA TTCCAGGGGAGAGTTACCCTCGAGCAAGAGCACCGGCGGCAGCGAAATTGTCCTGACTCA ACTGCCGATAAGTCCACATCAGTCTCCAGCCACACTGAGTGCATCTCCCGGCGAGCGGGTC ACCGCCTATATGGAATTGAGTACTCTTAGTTGCAGCGCCAGTTCTAGTGTATCATATATGAA TCCCTTCGTAGTGAGGACACTCTGGTATCAGCAAAAGCCAGGTCAAGCTCCCAGGCGATGG GCCGTCTACTACTGTGCCTCCCATATACGACTCATCAAAACTCGCCTCTGGCGTCCCAGCCCG CTCAGGTTCATTATGATTACTCGTTCTCCGGTTCCGGCTCTGGGCGCGACTATACCCTTACAA AGGTTTTCCATACTGGGGCCATTTCTAGCCTCGAACCAGAAGATTTTGCTGTATATTATTGT GGGCACCCTCGTAACAGTAAGCAACAGTGGTCACGTAACCCACCAACCTTCGGTGGAGGGA CAGCGGCGGCTCCGAGGGCACAAAGGTCGAGATAAAAGAGCCCAAATCTAGCGACAAAA AGAGCAGCGGCAGCGGCAGCCTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGG GAGAGCAAGAGCACCGGCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACA CAGCGAAATTGTCCTGACTCACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT GTCTCCAGCCACACTGAGTGCGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA ATCTCCCGGCGAGCGGGTCACCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC TCTTAGTTGCAGCGCCAGTTCTAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT AGTGTATCATATATGAACTGGGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT TATCAGCAAAAGCCAGGTCAAGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC GCTCCCAGGCGATGGATATACCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA GACTCATCAAAACTCGCCTCTGGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG GCGTCCCAGCCCGGTTCTCCGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GTTCCGGCTCTGGGCGCGACTGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG ATACCCTTACAATTTCTAGCCTAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC CGAACCAGAAGATTTTGCTGTCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC ATATTATTGTCAACAGTGGTCAAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC CGTAACCCACCAACCTTCGGTGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GGAGGGACAAAGGTCGAGATCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AAAAG (206) (207) CD3B2030-CAGGTGCAACTGGTACAAAGTGGTGCCGAGGTGAAGAAG CAGGTGCAACTGGTACAAAGT N106G-CCAGGGTCCAGTGTGAAAGTATCATGTAAAGCCAGCGGGT GGTGCCGAGGTGAAGAAGCC HL-scFvACACATTCACTAGGAGCACTATGCACTGGGTAAAGCAAGC AGGGTCCAGTGTGAAAGTATCCCCAGGGCAAGGTTTGGAGTGGATCGGTTATATTAACCCT ATGTAAAGCCAGCGGGTACACTCATCTGCTTATACAAATTACAATCAGAAATTCCAAGGGAG ATTCACTAGGAGCACTATGCAGGTCACTTTGACCGCTGACAAGTCTACCTCTACTGCATACA CTGGGTAAAGCAAGCCCCAGGTGGAACTCTCCAGCCTTCGTTCAGAAGACACAGCCGTTTAT GCAAGGTTTGGAGTGGATCGTACTGTGCCTCCCCACAGGTACACTACGACTACGGTGGATT GTTATATTAACCCTTCATCTGCCCCATATTGGGGTCAAGGCACCCTTGTAACAGTATCAAGC TTATACAAATTACAATCAGAAGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGA ATTCCAAGGGAGGGTCACTTTGAGCAAGAGCACCGGCGGCAGCGAAATTGTACTCACACA GACCGCTGACAAGTCTACCTCAAGTCCTGCAACTTTGTCTGCCTCACCAGGGGAAAGAGTA TACTGCATACATGGAACTCTCCACTCTTAGTTGTAGTGCTAGTTCATCCGTTTCTTATATGAAT AGCCTTCGTTCAGAAGACACATGGTATCAGCAGAAACCCGGACAAGCACCCCGGCGGTGG GCCGTTTATTACTGTGCCTCCCATATACGATTCCAGTAAACTTGCAAGCGGAGTCCCCGCAC CACAGGTACACTACGACTACGGTTTCAGCGGCAGTGGCTCAGGCCGGGACTATACCCTGAC GTGGATTCCCATATTGGGGTCTATTTCCTCCTTGGAACCTGAGGATTTTGCTGTGTACTACT AAGGCACCCTTGTAACAGTATGTCAGCAATGGAGTAGAAATCCTCCCACCTTTGGAGGTGG CAAGCGGCGGCTCCGAGGGCCACTAAAGTAGAGATCAAAGAGCCCAAATCTAGCGACAAA AAGAGCAGCGGCAGCGGCAGACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG CGAGAGCAAGAGCACCGGCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC GCAGCGAAATTGTACTCACACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT AAAGTCCTGCAACTTTGTCTGCGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA CTCACCAGGGGAAAGAGTAACCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC TCTTAGTTGTAGTGCTAGTTCAAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT TCCGTTTCTTATATGAATTGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT ATCAGCAGAAACCCGGACAAGGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC CACCCCGGCGGTGGATATACGCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA ATTCCAGTAAACTTGCAAGCGGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG GAGTCCCCGCACGTTTCAGCGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GCAGTGGCTCAGGCCGGGACTGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG ATACCCTGACTATTTCCTCCTTAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC GGAACCTGAGGATTTTGCTGTCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC GTACTACTGTCAGCAATGGAGAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC TAGAAATCCTCCCACCTTTGGAGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GGTGGCACTAAAGTAGAGATCCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AAAG (208) (209) CD3B2030-CAAGTGCAACTCGTGCAAAGCGGGGCTGAAGTGAAGAAG CAAGTGCAACTCGTGCAAAGC N106A-CCTGGATCAAGCGTGAAGGTCAGTTGCAAAGCCTCTGGAT GGGGCTGAAGTGAAGAAGCC HL-scFvATACCTTCACTCGATCAACCATGCACTGGGTCAAGCAGGC TGGATCAAGCGTGAAGGTCAGCCCAGGGCAAGGGCTCGAATGGATAGGATATATTAACCCA TTGCAAAGCCTCTGGATATACAGTTCTGCCTACACTAACTATAATCAGAAGTTTCAAGGCCG CTTCACTCGATCAACCATGCACGGTAACACTTACAGCCGATAAGAGTACCTCAACAGCATAC TGGGTCAAGCAGGCCCCAGGATGGAACTTAGTTCTTTGCGGAGCGAGGATACCGCTGTGT GCAAGGGCTCGAATGGATAGATTACTGCGCTTCACCTCAGGTTCACTACGACTACGCTGGA GATATATTAACCCAAGTTCTGCTTTCCCTATTGGGGTCAGGGTACACTGGTTACAGTTTCCTC CTACACTAACTATAATCAGAATGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCG GTTTCAAGGCCGGGTAACACTAGAGCAAGAGCACCGGCGGLAGCGAAATTGTTTTGACCC TACAGCCGATAAGAGTACCTCAATCACCTGCCACTCTCTCTGCCTCTCCTGGTGAGCGAGTT AACAGCATACATGGAACTTAGACTTTGTCATGTAGCGCATCATCAAGTGTATCTTACATGAA TTCTTTGCGGAGCGAGGATACCTGGTACCAACAAAAACCCGGACAGGCACCACGTCGTTGG CGCTGTGTATTACTGCGCTTCAATTTATGACAGTAGCAAGCTCGCCTCCGGGGTACCCGCAA CCTCAGGTTCACTACGACTACGATTTTCCGGGTCAGGGTCTGGCAGGGACTATACCCTGAC GCTGGATTTCCCTATTGGGGTAATCAGCAGTCTGGAACCTGAGGACTTTGCTGTGTATTACT CAGGGTACACTGGTTACAGTTGCCAACAGTGGTCTCGCAACCCCCCTACTTTCGGGGGAGG TCCTCTGGCGGCTCCGAGGGCTACAAAGGTAGAAATTAAGGAGCCCAAATCTAGCGACAAA AAGAGCAGCGGCAGCGGCAGACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG CGAGAGCAAGAGCACCGGCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC GCAGCGAAATTGTTTTGACCCACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT AATCACCTGCCACTCTCTCTGCGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA CTCTCCTGGTGAGCGAGTTACCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC TTTGTCATGTAGCGCATCATCAAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT AGTGTATCTTACATGAACTGGGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT TACCAACAAAAACCCGGACAGGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC GCACCACGTCGTTGGATTTATCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA GACAGTAGCAAGCTCGCCTCCGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG GGGGTACCCGCAAGATTTTCCGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GGGTCAGGGTCTGGCAGGGAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG CTATACCCTGACAATCAGCAGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC TCTGGAACCTGAGGACTTTGCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC TGTGTATTACTGCCAACAGTGAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC GTCTCGCAACCCCCCTACTTTCGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA GGGGGAGGTACAAAGGTAGACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT AATTAAGG (210) (211) CD3B2030-CAAGTTCAGTTGGTTCAATCCGGCGCAGAGGTTAAAAAAC CAAGTTCAGTTGGTTCAATCC HL-scFvCCGGATCAAGCGTTAAGGTTAGTTGTAAAGCCTCTGGCTA GGCGCAGAGGTTAAAAAACCCCACTTTCACACGCTCAACAATGCATTGGGTTAAGCAGGCCC GGATCAAGCGTTAAGGTTAGTCTGGGCAGGGACTGGAGTGGATCGGTTACATAAACCCATC TGTAAAGCCTCTGGCTACACTTCAGCGCCTATACAAACTATAACCAGAAGTTCCAAGGGCGG TCACACGCTCAACAATGCATTGTTACATTGACCGCTGACAAGTCCACTAGCACAGCATATAT GGGTTAAGCAGGCCCCTGGGCGGAGCTGTCAAGTCTGAGATCCGAAGACACTGCCGTATAT AGGGACTGGAGTGGATCGGTTATTGCGCTAGTCCACAAGTGCACTATGACTATAACGGTTT TACATAAACCCATCCAGCGCCTTCCCTATTGGGGACAAGGAACCCTGGTGACCGTTAGCTCC ATACAAACTATAACCAGAAGTGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGA TCCAAGGGCGGGTTACATTGAGAGCAAGAGCACCGGCGGCAGCGAAATTGTCTTGACCCA CCGCTGACAAGTCCACTAGCAGTCTCCAGCAACTCTTAGTGCATCACCAGGTGAGCGTGTTA CAGCATATATGGAGCTGTCAACCCTCTCATGTAGCGCCAGCTCATCTGTTAGTTATATGAAT GTCTGAGATCCGAAGACACTGTGGTATCAACAGAAACCAGGGCAAGCTCCCAGAAGATGG CCGTATATTATTGCGCTAGTCCATATATGATTCTTCAAAACTCGCAAGTGGTGTCCCAGCCCG ACAAGTGCACTATGACTATAACTTCTCAGGCTCTGGTTCCGGTCGCGATTATACTCTCACCA CGGTTTTCCCTATTGGGGACATCAGTAGTTTGGAACCCGAAGATTTCGCCGTCTATTATTGC AGGAACCCTGGTGACCGTTAGCAGCAATGGAGCAGGAATCCCCCCACATTCGGCGGCGGTA CTCCGGCGGCTCCGAGGGCAACAAAGGTTGAGATTAAGGAGCCCAAATCTAGCGACAAAAC GAGCAGCGGCAGCGGCAGCGTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGG AGAGCAAGAGCACCGGCGGCGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACAC AGCGAAATTGTCTTGACCCAGCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTG TCTCCAGCAACTCTTAGTGCATGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACT CACCAGGTGAGCGTGTTACCCGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAA TCTCATGTAGCGCCAGCTCATCAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGG TGTTAGTTATATGAATTGGTATTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGG CAACAGAAACCAGGGCAAGCTCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCA CCCAGAAGATGGATATATGATGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGC TCTTCAAAACTCGCAAGTGGTCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGGGA GTCCCAGCCCGCTTCTCAGGCTGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTC CTGGTTCCGGTCGCGATTATAAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGA CTCTCACCATCAGTAGTTTGGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTC ACCCGAAGATTTCGCCGTCTATCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGCAA TATTGCCAGCAATGGAGCAGGGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGT AATCCCCCCACATTCGGCGGCCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACT GGTACAAAGGTTGAGATTAAGACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT G (212) (213) CD3B2030-GAGATCGTTCTGACACAGTCTCCCGCAACCCTCAGCGCTTC GAGATCGTTCTGACACAGTCT N106Q-ACCCGGTGAGCGTGTCACTCTGAGCTGTTCCGCTAGTAGT CCCGCAACCCTCAGCGCTTCAC LH-scFvAGCGTTAGCTACATGAACTGGTATCAACAAAAGCCAGGAC CCGGTGAGCGTGTCACTCTGAAGGCACCCAGGCGATGGATTTACGATTCATCAAAACTGGC GCTGTTCCGCTAGTAGTAGCGAAGCGGAGTGCCTGCTCGTTTTAGTGGGTCCGGGTCTGGC TTAGCTACATGAACTGGTATCCGCGATTACACCCTGACCATATCATCCCTCGAACCTGAGGA AACAAAAGCCAGGACAGGCACTTCGCAGTTTATTATTGCCAACAGTGGAGTAGGAACCCAC CCCAGGCGATGGATTTACGATCTACATTCGGTGGGGGGACCAAAGTCGAGATAAAAGGCG TCATCAAAACTGGCAAGCGGAGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGC GTGCCTGCTCGTTTTAGTGGGAAGAGCACCGGCGGCAGCCAGGTACAACTCGTACAGAGC TCCGGGTCTGGCCGCGATTACGGAGCTGAGGTGAAAAAACCCGGTAGTTCCGTTAAGGTTA ACCCTGACCATATCATCCCTCGGCTGTAAGGCTAGCGGATACACTTTTACTCGATCTACAATG AACCTGAGGACTTCGCAGTTTCACTGGGTTAAACAGGCTCCCGGCCAGGGTTTGGAATGGA ATTATTGCCAACAGTGGAGTATCGGATACATCAACCCCAGTAGTGCCTATACCAATTACAAT GGAACCCACCTACATTCGGTGCAAAAGTTTCAAGGCAGAGTGACCCTGACCGCTGACAAAT GGGGGACCAAAGTCGAGATACCACAAGTACCGCATATATGGAGCTCTCAAGTTTGCGAAG AAAGGCGGCTCCGAGGGCAATGAAGATACTGCTGTATATTATTGCGCAAGCCCTCAAGTTC GAGCAGCGGCAGCGGCAGCGACTATGACTATCAAGGGTTTCCTTACTGGGGTCAGGGAAC AGAGCAAGAGCACCGGCGGCACTGGTCACAGTATCATCCGAGCCCAAATCTAGCGACAAA AGCCAGGTACAACTCGTACAGACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAG AGCGGAGCTGAGGTGAAAAAGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC ACCCGGTAGTTCCGTTAAGGTACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGT TAGCTGTAAGGCTAGCGGATAGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAA CACTTTTACTCGATCTACAATGCTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGAC CACTGGGTTAAACAGGCTCCCAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT GGCCAGGGTTTGGAATGGATCGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT GGATACATCAACCCCAGTAGTGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC GCCTATACCAATTACAATCAAACAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCA AGTTTCAAGGCAGAGTGACCCGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATCCCGG TGACCGCTGACAAATCCACAAGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTG GTACCGCATATATGGAGCTCTGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGG CAAGTTTGCGAAGTGAAGATAAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGC CTGCTGTATATTATTGCGCAAGCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGTGAGC CCCTCAAGTTCACTATGACTATAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAAC CAAGGGTTTCCTTACTGGGGTGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCA CAGGGAACACTGGTCACAGTACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT TCATCCG (214) (215) CD3B2030-CAGGTACAACTCGTACAGAGCGGAGCTGAGGTGAAAAAA CAGGTACAACTCGTACAGAGC N106Q-CCCGGTAGTTCCGTTAAGGTTAGCTGTAAGGCTAGCGGAT GGAGCTGAGGTGAAAAAACC HL-scFvACACTTTTACTCGATCTACAATGCACTGGGTTAAACAGGCT CGGTAGTTCCGTTAAGGTTAGCCCGGCCAGGGTTTGGAATGGATCGGATACATCAACCCCA CTGTAAGGCTAGCGGATACACGTAGTGCCTATACCAATTACAATCAAAAGTTTCAAGGCAG TTTTACTCGATCTACAATGCACAGTGACCCTGACCGCTGACAAATCCACAAGTACCGCATAT TGGGTTAAACAGGCTCCCGGCATGGAGCTCTCAAGTTTGCGAAGTGAAGATACTGCTGTAT CAGGGTTTGGAATGGATCGGAATTATTGCGCAAGCCCTCAAGTTCACTATGACTATCAAGGG TACATCAACCCCAGTAGTGCCTTTTCCTTACTGGGGTCAGGGAACACTGGTCACAGTATCATC ATACCAATTACAATCAAAAGTTCGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCG TCAAGGCAGAGTGACCCTGACAGAGCAAGAGCACCGGCGGCAGCGAGATCGTTCTGACAC CGCTGACAAATCCACAAGTACAGTCTCCCGCAACCCTCAGCGCTTCACCCGGTGAGCGTGTC CGCATATATGGAGCTCTCAAGACTCTGAGCTGTTCCGCTAGTAGTAGCGTTAGCTACATGAA TTTGCGAAGTGAAGATACTGCCTGGTATCAACAAAAGCCAGGACAGGCACCCAGGCGATG TGTATATTATTGCGCAAGCCCTGATTTACGATTCATCAAAACTGGCAAGCGGAGTGCCTGCT CAAGTTCACTATGACTATCAACGTTTTAGTGGGTCCGGGTCTGGCCGCGATTACACCCTGA GGGTTTCCTTACTGGGGTCAGCCATATCATCCCTCGAACCTGAGGACTTCGCAGTTTATTAT GGAACACTGGTCACAGTATCATGCCAACAGTGGAGTAGGAACCCACCTACATTCGGTGGGG TCCGGCGGCTCCGAGGGCAAGGACCAAAGTCGAGATAAAAGAGCCCAAATCTAGCGACA GAGCAGCGGCAGCGGCAGCGAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGC AGAGCAAGAGCACCGGCGGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAG AGCGAGATCGTTCTGACACAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGT TCTCCCGCAACCCTCAGCGCTTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTT CACCCGGTGAGCGTGTCACTCCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAA TGAGCTGTTCCGCTAGTAGTAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCG GCGTTAGCTACATGAACTGGTTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTG ATCAACAAAAGCCAGGACAGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCC GCACCCAGGCGATGGATTTACCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAG GATTCATCAAAACTGGCAAGCGGCAGCCCCGAGAACCACAGGTGTACGTGTACCCCCCATC GGAGTGCCTGCTCGTTTTAGTCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTG GGGTCCGGGTCTGGCCGCGATCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAG TACACCCTGACCATATCATCCCTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACC TCGAACCTGAGGACTTCGCAGACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCGCCCTCGT TTTATTATTGCCAACAGTGGAGAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGG GTAGGAACCCACCTACATTCGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACA GTGGGGGGACCAAAGTCGAGACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT ATAAAAG (216) (217) CD3B2029-CAGGTGCAGCTGGTGCAGAGCGGCGCGGAAGTGAAAAAA CAGGTGCAGCTGGTGCAGAG N106QCCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCGAGCGGC CGGCGCGGAAGTGAAAAAAC HL scFvTATACCTTTACCCGCAGCACCATGCATTGGGTGAAACAGG CGGGCAGCAGCGTGAAAGTGCGCCGGGCCAGGGCCTGGAATGGATTGGCTATATTAACCC AGCTGCAAAGCGAGCGGCTATGAGCAGCGCGTATACCAACTATAACCAGAAATTTCAGGGC ACCTTTACCCGCAGCACCATGCCGCGTGACCCTGACCGCGGATAAAAGCACCAGCACCGCGT ATTGGGTGAAACAGGCGCCGATATGGAACTGAGCAGCCTGCGCAGCGAAGATACCGCGG GGCCAGGGCCTGGAATGGATTTGTATTATTGCGCGAGCCCGCAGGTGCATTATGATTATCA GGCTATATTAACCCGAGCAGCGGGCTTTCCGTATTGGGGCCAGGGCACCCTGGTGACCGTG GCGTATACCAACTATAACCAGAGCAGCGGCGGCAGCGAAGGCAAAAGCAGCGGCAGCGG AAATTTCAGGGCCGCGTGACCCAGCGAAAGCAAAAGCACCGGCGGCAGCGAAATTGTGCT CTGACCGCGGATAAAAGCACCGACCCAGAGCCCGGCGACCCTGAGCGCGAGCCCGGGCGA AGCACCGCGTATATGGAACTGACGCGTGACCCTGAGCTGCAGCGCGAGCAGCAGCGTGAG AGCAGCCTGCGCAGCGAAGATCTATATGAACTGGTATCAGCAGAAACCGGGCCAGAGCCCG ACCGCGGTGTATTATTGCGCGCGCCGCTGGATTTATGATAGCAGCAAACTGGCGAGCGGC AGCCCGCAGGTGCATTATGATGTGCCGGCGCGCTTTAGCGGCAGCGGCAGCGGCCGCGAT TATCAGGGCTTTCCGTATTGGTATACCCTGACCATTAGCAGCCTGGAACCGGAAGATTTTG GGCCAGGGCACCCTGGTGACCCGGTGTATTATTGCCAGCAGTGGAGCCGCAACCCGCCGAC GTGAGCAGCGGCGGCAGCGACTTTGGCGGCGGCACCAAAGTGGAAATTAAAGAACCGAA AGGCAAAAGCAGCGGCAGCGAAGCAGCGATAAAACCCATACCTGCCCGCCGTGCCCGGCG GCAGCGAAAGCAAAAGCACCCCGGAAGCGGCGGGCGGCCCGAGCGTGTTTCTGTTTCCGC GGCGGCAGCGAAATTGTGCTGCGAAACCGAAAGATACCCTGATGATTAGCCGCACCCCGGA ACCCAGAGCCCGGCGACCCTGAGTGACCTGCGTGGTGGTGAGCGTGAGCCATGAAGATCC AGCGCGAGCCCGGGCGAACGGGAAGTGAAATTTAACTGGTATGTGGATGGCGTGGAAGT CGTGACCCTGAGCTGCAGCGCGCATAACGCGAAAACCAAACCGCGCGAAGAACAGTATAA GAGCAGCAGCGTGAGCTATATCAGCACCTATCGCGTGGTGAGCGTGCTGACCGTGCTGCAT GAACTGGTATCAGCAGAAACCCAGGATTGGCTGAACGGCAAAGAATATAAATGCAAAGTG GGGCCAGAGCCCGCGCCGCTAGCAACAAAGCGCTGCCGGCGCCGATTGAAAAAACCATTA GGATTTATGATAGCAGCAAACGCAAAGCGAAAGGCCAGCCGCGCGAACCGCAGGTGTATG TGGCGAGCGGCGTGCCGGCGTGTATCCGCCGAGCCGCGAAGAAATGACCAAAAACCAGGT CGCTTTAGCGGCAGCGGCAGCGAGCCTGACCTGCCTGGTGAAAGGCTTTTATCCGAGCGAT GGCCGCGATTATACCCTGACCATTGCGGTGGAATGGGAAAGCAACGGCCAGCCGGAAAAC ATTAGCAGCCTGGAACCGGAAAACTATAAAACCACCCCGCCGGTGCTGGATAGCGATGGCA GATTTTGCGGTGTATTATTGCCGCTTTGCGCTGGTGAGCAAACTGACCGTGGATAAAAGCCG AGCAGTGGAGCCGCAACCCGCCTGGCAGCAGGGCAACGTGTTTAGCTGCAGCGTGATGCAT CGACCTTTGGCGGCGGCACCAGAAGCGCTGCATAACCATTATACCCAGAAAAGCCTGAGCC AAGTGGAAATTAAA TGAGCCCGGGC(219) (218)

Engineering of CD3 Fabs for BCMAxCD3 Bispecific Generation

The CD3 specific VH and VL regions were engineered inVH-CH1-linker-CH2-CH3 and VL-CL formats respectively and expressed asIgG1. The polypeptide of SEQ ID NO: 220 comprising the Fc silencingmutation L234A/L235A/D265S and the CH3 mutation T350V/L351Y/F405A/Y407Vdesigned to promote selective heterodimerization was used to generatethe CD3 specific VH-CH1-linker-CH2-CH3 (Table 25).

(huIgG1_G1m(17) _AAS_ZWA) SEQ ID NO: 220ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPR EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSREEMTKNQ VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGKThe polypeptides of SEQ ID NO: 221 or 222 were used to generate the CD3specific VL-CL (Table 26)

(human kappa light chain) SEQ ID NO: 221RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG EC (human lambda light chain)SEQ ID NO: 222 GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSDNA sequences of anti-CD3 molecules as HC in VH-CH1-liker-CH2-CH3 formatand LC in VL-CL format are shown in Table 27.

TABLE 25 Amino acid sequence of the anti-CD3 antibodyarm VH-CH1-linker-CH2-CH3 of the bi- specific antibody. SEQ HC IDprotein BsAb NO: HC amino acid sequence Cris7b GCDB131 223QVQLLQSAAEVKKPGESLKI SCKGSGYTFTRSTMHWVRQT PGKGLEWMGYINPSSAYTNYNQKFKDQVTISADKSISTAY LQWSSLKASDTAMYYCARPQ VHYDYNGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVE PKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRT PEVTCVVVSVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYVYPPSREEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPP VLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPG Cris7b- HC3B127 224 QVQLLQSAAEVKKPGESLKIN106Q SCKGSGYTFTRSTMHWVRQT PGKGLEWMGYINPSSAYTNY NQKFKDQVTISADKSISTAYLQWSSLKASDTAMYYCARPQ VHYDYQGFPYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVE PKSCDKTHTCPPCPAPEAAG GPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFN WYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYVYPPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFALVSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPG

TABLE 26 Amino acid sequence of the anti-CD3 antibodylight chain arm (VL-CL) of the bi-specific antibody LC SEQ protein BsAbID NO: LC amino acid sequence Cris7b GCDB 225 EIVLTQSPSAMSASVGDRVT 131ITCSASSSVSYMNWYQQKPG KVPKRLIYDSSKLASGVPSR FSGSGSGTEYTLTISSLQPEDFATYYCQQWSRNPPTFGQG TMLEIKRTVAAPSVFIFPPS DEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Cris7b- HC3B1 226 EIVLTQSPSAMSASVGDRVT N106Q 27ITCSASSSVSYMNWYQQKPG KVPKRLIYDSSKLASGVPSR FSGSGSGTEYTLTISSLQPEDFATYYCQQWSRNPPTFGQG TMLEIKRTVAAPSVFIFPPS DEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

TABLE 27 cDNA SEQ ID NOs of anti-CD3 ars of bi-specificantibodies HC in VH-CHl-liker-CH2- CH3 format and LC in VL-CL format.HC cDNA LC cDNA ID BsAb SEQ ID NO: SEQ ID NO: Cris7b GCDB131caggtgcagctgctg gaaattgtgctgacc cagagcgcggcggaa cagagcccgagcgcggtgaaaaaaccgggc atgagcgcgagcgtg gaaagcctgaaaatt ggcgatcgcgtgaccagctgcaaaggcagc attacctgcagcgcg ggctatacctttacc agcagcagcgtgagccgcagcaccatgcat tatatgaactggtat tgggtgcgccagacc cagcagaaaccgggcccgggcaaaggcctg aaagtgccgaaacgc gaatggatgggctat ctgatttatgatagcattaacccgagcagc agcaaactggcgagc gcgtataccaactat ggcgtgccgagccgcaaccagaaatttaaa tttagcggcagcggc gatcaggtgaccatt agcggcaccgaatatagcgcggataaaagc accctgaccattagc attagcaccgcgtat agcctgcagccggaactgcagtggagcagc gattttgcgacctat ctgaaagcgagcgat tattgccagcagtggaccgcgatgtattat agccgcaacccgccg tgcgcgcgcccgcag acctttggccagggcgtgcattatgattat accatgctggaaatt aacggctttccgtat aaacgtacggtggcttggggccagggcacc gcaccatctgtcttc ctggtgaccgtgagc atcttcccgccatctagcgcctccaccaag gatgagcagttgaaa ggcccatcggtcttc tctggaactgectctcccctggcaccctcc gttgtgtgcctgctg tccaagagcacctct aataacttctatcccgggggcacagcggcc agagaggccaaagta ctgggctgcctggtc cagtggaaggtggataaggactacttcccc aacgccctccaatcg gaaccggtgacggtg ggtaactcccaggagtcgtggaactcaggc agtgtcacagagcag gccctgaccagcggc gacagcaaggacagcgtgcacaccttcccg acctacagcctcagc gctgtcctacagtcc agcaccctgacgctgtcaggactctactcc agcaaagcagactac ctcagcagcgtggtg gagaaacacaaagtcaccgtgccctccagc tacgcctgcgaagtc agcttgggcacccag acccatcagggcctgacctacatctgcaac agctcgcccgtcaca gtgaatcacaagccc aagagcttcaacaggagcaacaccaaggtg ggagagtgt gacaagaaagttgag (228) cccaaatcttgtgacaaaactcacacatgt ccaccgtgcccagca cctgaagcagcaggg ggaccgtcagtcttcctcttccccccaaaa cccaaggacaccctc atgatctcccggacc cctgaggtcacatgcgtggtggtgagcgtg agccacgaagaccct gaggtcaagttcaac tggtacgtggacggcgtggaggtgcataat gccaagacaaagccg cgggaggagcagtac aacagcacgtaccgtgtggtcagcgtcctc accgtcctgcaccag gactggctgaatggc aaggagtacaagtgcaaggtctccaacaaa gccctcccagccccc atcgagaaaaccatc tccaaagccaaagggcagccccgagaacca caggtgtacgtgtac cccccatcccgggag gagatgaccaagaaccaggtcagcctgacc tgcctggtcaaaggc ttctatcccagcgac atcgccgtggagtgggagagcaatgggcag ccggagaacaactac aagaccacgcctccc gtgctggactccgacggctccttcgccctc gtgagcaagctcacc gtggacaagtctaga tggcagcaggggaacgtcttctcatgctcc gtgatgcatgaggct ctgcacaaccactac acgcagaagagcctctccctgtctccgggt (227) Cris7b- HC3B127 caagtgcaactcctt gaaattgtgctgaccN106Q cagtcagccgccgag cagagcccgagcgcg gttaaaaaaccagga atgagcgcgagcgtggaatcactgaaaatc ggcgatcgcgtgacc tcctgtaagggtagc attacctgcagcgcgggatataccttcact agcagcagcgtgagc agatcaaccatgcat tatatgaactggtattgggtgagacagact cagcagaaaccgggc ccaggtaaaggattg aaagtgccgaaacgcgagtggatgggatac ctgatttatgatagc ataaacccctcctca agcaaactggcgagcgcctataccaattac ggcgtgccgagccgc aatcaaaaatttaag tttagcggcagcggcgatcaagtgactatc agcggcaccgaatat agtgctgacaagagc accctgaccattagcatctcaaccgcctac agcctgcagccggaa cttcagtggtcatca gattttgcgacctatctgaaagcatcagat tattgccagcagtgg acagccatgtattac agccgcaacccgccgtgtgcaagaccccaa acctttggccagggc gttcactatgactat accatgctggaaattcagggtttcccatac aaacgtacggtggct tgggggcaaggaaca gcaccatctgtcttcctcgtgaccgtttca atcttcccgccatct tctgcctccaccaag gatgagcagttgaaaggcccatcggtcttc tctggaactgectct cccctggcaccctcc gttgtgtgcctgctgtccaagagcacctct aataacttctatccc gggggcacagcggcc agagaggccaaagtactgggctgcctggtc cagtggaaggtggat aaggactacttcccc aacgccctccaatcggaaccggtgacggtg ggtaactcccaggag tcgtggaactcaggc agtgtcacagagcaggccctgaccagcggc gacagcaaggacagc gtgcacaccttcccg acctacagcctcagcgctgtcctacagtcc agcaccctgacgctg tcaggactctactcc agcaaagcagactacctcagcagcgtggtg gagaaacacaaagtc accgtgccctccagc tacgcctgcgaagtcagcttgggcacccag acccatcagggcctg acctacatctgcaac agctcgcccgtcacagtgaatcacaagccc aagagcttcaacagg agcaacaccaaggtg ggagagtgtgacaagaaagttgag (228) cccaaatcttgtgac aaaactcacacatgt ccaccgtgcccagcacctgaagcagcaggg ggaccgtcagtcttc ctcttccccccaaaa cccaaggacaccctcatgatctcccggacc cctgaggtcacatgc gtggtggtgagcgtg agccacgaagaccctgaggtcaagttcaac tggtacgtggacggc gtggaggtgcataat gccaagacaaagccgcgggaggagcagtac aacagcacgtaccgt gtggtcagcgtcctc accgtcctgcaccaggactggctgaatggc aaggagtacaagtgc aaggtctccaacaaa gccctcccagcccccatcgagaaaaccatc tccaaagccaaaggg cagccccgagaacca caggtgtacgtgtaccccccatcccgggag gagatgaccaagaac caggtcagcctgacc tgcctggtcaaaggcttctatcccagcgac atcgccgtggagtgg gagagcaatgggcag ccggagaacaactacaagaccacgcctccc gtgctggactccgac ggctccttcgccctc gtgagcaagctcaccgtggacaagtctaga tggcagcaggggaac gtcttctcatgctcc gtgatgcatgaggctctgcacaaccactac acgcagaagagcctc tccctgtctccgggt (229)

Engineering of BCMA Fab-Fc for BCMAxCD3 Bispecific Generation

The BCMA specific VH and VL regions were engineered inVH-CH1-linker-CH2-CH3 and VL-CL formats respectively. The polypeptide ofSEQ ID NO: 230 comprising the Fc silencing mutation L234A/L235A/D265Sand the CH3 mutation T350V/T366L/K392L/T394W designed to promoteselective heterodimerization was used to generate the CD3 specificVH-CH1-linker-CH2-CH3).

(huIgG1_G1m(17)_AAS_ZWB) SEQ ID NO: 230ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPR EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSREEMTKNQ VSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGKThe polypeptides of SEQ ID NO: 231 or 232 were used to generate the BCMAspecific VL-CL.

(human kappa light chain) SEQ ID NO: 231RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG EC (human lambda light chain)SEQ ID NO: 232 GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSThe amino acid sequences of BCMA Fab-Fc heavy chain (HC) and lightchains (LCs) are shown below.

BCMA Fab-Fc heavy chain (SEQ ID NO: 233)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMS WVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDEGYSSG HYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHT FPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPS VFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPSDIAV EWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG BCMA Fab-Fc light chain(SEQ ID NO: 234) EIVLTQSPGTLSLSPGERATLSCRASQSISSSFLTWYQQKPGQAPRLLIYGASSRATGIPDRFSGGGSGT DFTLTISRLEPEDFAVYYCQHYGSSPMYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKS FNRGECEngineering of BCMA scFvs-Fc for BCMA×CD3 Bispecific Generation

BCMA VH/VL regions engineered as scFvs in either VH-Linker-VL orVL-linker-VH orientations using the linker of SEQ ID NO: 3 (Table 2), asdescribed in Example 2, were further engineered into ascFv-hinge-CH2-CH3 format comprising the Fc silencing mutation(L234A/L235A/D265S) and the T350V/T366L/K392L/T394W mutations designedto promote selective heterodimerization and expressed as IgG1 (Table28). The polypeptide of SEQ ID NO: 235 was used as the constant domainhinge-CH2-CH3 (Fc).

SEQ ID NO: 235 (huIgG1_G1m(17)-hinge-Fc_C220S_AAS_ZWB)EPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDT LMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPSDIAVEWESNGQPENN YLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

TABLE 28 Amino acid sequences of anti- BCMA scFvs-Fcfor BCMAxCD3 bispecific generation Protein SEQ ID NO:Amino acid sequence BCMB519-LH-scFv 236 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQA PGKGLEWVSAISGSGGSTYY ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDE GYSSGHYYGMDVWGQGTTVT VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSCDKTHTCPPCPAPEA AGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVK FNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYVLPPS REEMTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTW PPVLDSDGSFFLYSKLTVDK SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

The bsAbs were assayed for thermal stability, abilities to bind T cells,and cytotoxicity.

Thermal Stability Analysis

For thermal stability, Thermal unfolding and aggregation were measuredfrom 20° C.-95° C. using a ramp of 1° C./min on a NanoTemperPromethiusNT.48 instrument. Samples were then transferred into nanoDSFcapillary by capillary action and assayed in duplicate (Table 2). Allvariants (except for BC3B126) had similar Tonset, Tm1, and Tagg,indicating that neither the specific back-mutations nor the identity ofthe mutation at VH N106 had a significant impact on thermostability. Onaverage the bsAbs had Tonset, Tm1, and Tagg of 61, 68, and 77° C., whereTm1 represented the melting temperature of the scFv moiety, suggestingthat all the Cris-7 variants could be amenable to therapeuticdevelopment.

TABLE Ther mostability analysis of bsAbs containing Cris-7 scFvmoieties. Tm/Tagg Protein Batch Name Tonset Tml Tagg BC3B51.001 61.8° C.69.1° C. 77.7° C. BC3B109.001 61.0° C. 68.3° C. 76.3° C. BC3B114.00161.1° C. 68.4° C. 78.2° C. BC3B53.001 61.4° C. 68.9° C. 79.2° C.BC3B123.001 62.0° C. 68.8° C. 76.8° C. BC3B128.001 60.7° C. 68.7° C.77.5° C. BC3B103.001 61.4° C. 68.8° C. 78.2° C. BC3B107.001 61.4° C.68.2° C. 77.6° C. BC3B112.001 NA NA NA BC3B117.001 61.4° C. 69.1° C.77.7° C. BC3B121.001 61.7° C. 68.3° C. 76.6° C. BC3B126.001 51.1° C.68.1° C. 76.5° C. BC3B104.001 60.5° C. 69.0° C. 76.5° C. BC3B108.00161.4° C. 68.1° C. 77.7° C. BC3B113.001 61.1° C. 68.2° C. 77.2° C.BC3B118.001 62.0° C. 69.2° C. 77.3° C. BC3B122.001 61.3° C. 68.5° C.76.3° C. BC3B127.001 61.4° C. 68.6° C. 75.9° C. BC3B105.001 61.8° C.69.3° C. 77.9° C. BC3B110.001 61.7° C. 68.0° C. 77.3° C. BC3B115.00161.0° C. 68.8° C. 76.5° C. BC3B119.001 61.7° C. 69.6° C. 77.8° C.BC3B124.001 NA NA NA BC3B129.001 61.4° C. 69.3° C. 77.2° C.

Activity Analysis

The BCMAxCD3 bi-specifics (BsAbs) were tested for their abilities toeither bind T cells or to induce T cell-based cytotoxicity against cellsexpressing each antigen (Tables 29 and 30). Binding and cytotoxicityassays are described below.

The bsAbs were assayed for their abilities bind T cells and to induceT-cell mediated cytotoxicity against H929 cells (ATCC® CRL-9068™). For Tcell binding, briefly, bsAbs were prepared at 2× concentration of 600 nMin assay media (RPMI 1640+10% HI-FBS) and diluted in 3-fold serialdilutions in sterile polypropylene (PP) greiner plates in stain bufferfor a 11-point titration. Stain buffer with no antibody was added tobottom 4 wells and assign as background control (secondary antibodyalone). Parental Cris7b, CD3B695 (bivalent mAb controls) and CD3B375(bsAb which was monovalent for Cris-7b) were added also with fulldose-response curves.

Frozen T-cells were thawed in a 37° C. water bath and transferred gentlyinto conical tube containing warm 5 mL media (RPMI+10% FBS)/1 vial of1×10⁶ cells. Cells were mixed and centrifuged for 5 minutes at 400×gfollowed by resuspension in flow staining buffer, and counted andviability checked. Cells were then plated in 50 uL/50,000 cells/wellinto assay plates. Assay positive control mAb was added in the firstcolumn in quadruplicates at 2× at 20 nM in the bottom 4 wells. Stainbuffer with no mAb was added to the top 4 wells and assigned asbackground controls (secondary antibody alone). Serially dilutedantibody samples were added at 50 uL/well using Intergra Viafloaccording to the attached plate maps and incubated for 1 hr at 37° C.After one hour incubation, 150 uL staining buffer were added to allwells, and cells were spun at 500×g for 5 minutes to pellet cells. Cellswere then washed prior to addition of Alexa Fluor 647 (A647) conjugatedanti human IgG Fc specific secondary detection antibody at 2 ug/mL instaining buffer. Secondary detection antibody was added at 50 uL/well tothe washed cells. Plates were covered with foil and incubated for 30minutes on ice or in the fridge. 150 uL staining buffer were added toall wells, and cells were spun at 500×g for 5 min to pellet cells. Cellswere resuspended in 20 uL running buffer containing 1:1000 dilution ofSYTOX™ green dead cell stain and run plates on iQue® Screener flowcytometer (Essen BioScience, Inc.). Briefly, cells were gated on FCS vs.SCS dot plot to eliminate debris. Singlets were gated on SCS-A vs SCS-Hdot plot and from singlet population, live cells gated choosing BL1channel for low/negative with SYTOX™ green viability stain (ThermoFisher). Cell binding of control mAbs or test panel supernatants wasassessed by comparing to negative/isotype control binding by RL1 (A647)Geomeans from the live cell population.

Antibody Sample Preparation

BsAbs were prepared at 20 nM in assay media and serially diluted 3-foldfor an 11-point titration in assay media and stored at 4 C.

Target cells (H929, ATCC® CRL-9068™) were prepared by addition of Fcblock at 5 uL/1×10⁶ cells (50 uL per 10 million cells), and incubated atroom temperature for 20 minutes at 37° C. Cells were diluted to 4×10⁵cells/ml for plating. Control wells were supplemented with 50 uL ofmedia/well and incubated at 37° C. T cell vials were thawed in at 37° C.into a 50 mL conical tube containing room temperature assay media (5 mLmedia/1 vial of cells). Cells were spun at 300×g for 5 min, andresuspended in 10 mL fresh media and counted. For E:T ratio of 5:1, aT-cell suspension at 2×10{circumflex over ( )}6/mL was prepared. Cellswere prepared at 50 uL/100 k/well to the assay plates already containingtumor target cells (from step above) according to the plate map. Controlwells were supplemented with 50 uL of media/well and incubated at 37° C.

Antibodies were added at 100 uL/well and serially diluted starting from10 nM to the assay plates containing mixed tumor target cells andT-cells. Target cells were at 20,000 cells/well and pan T-cells cellcounts at 100,000 cells/well. Total assay volume was 200 uL/well. Plateswere incubated at 37° C., 5% CO2 in a humidified cell culture incubatorfor 48 hr.

Cells were washed by adding 150 uL of BD staining buffer and spun at300×g for 5 min. Staining solution mixture in BD stain buffer contained:

APC-conjugated anti-Human CD4 (1:100) (R&D Systems FAB3791A-025)

APC-conjugated anti-Human CD8 (1:100) (R&D Systems FAB1509A-025)

Brilliant Violet 421™-conjugated anti-Human CD25 (1:500)

Vybrant™ DyeCycle™ Green Stain (Invitrogen) at 1:12500 dilution wereprepared separately and added at 50 uL/well staining solution mixture toall wells of assay plates. 25 uL/well of diluted vibrant green dye wereadded to top 2 wells of 1st control column of all assay plates followedby incubation at room temperature for 20 min. Plates were washed 2× with150 uL staining buffer and resuspended in 30 uL IntelliCyt® runningbuffer containing SYTOX™ green live/dead stain (1:1000) and analyzedusing the iQue® PLUS Screener within 4 hr.

Cells were gated on FSC-H vs SSC-H, and T-cell and tumor cellpopulations were gated from all cells on on APC (RL1) vs SSC-H. Livecells and dead cells (live/dead stain) were gated for both tumor and Tcells from their respective dot plots on FSC-H vs Sytox Green (BL2).Using live T-cells, activated/CD25 positive T cell populations weregated on FSC-H vs Brilliant Violet (VL1). Cell populations weredetermined as follows:

Dead tumor cells=SytoxGreen live-dead stain positive/total tumorcells×100

% Live T-cells=L/D negative T-cells/total T-cells×100

% Activated T-cells=CD25 positive Live T-cells/Live T-cells×100

Cell binding analysis showed that the Cris-7 variants displayed a rangeof affinity for T cells, and this cell-based affinity was correlated tothe EC50 for cytotoxicity (Table 29). In general, variants of the Cris-7v-region formatted in heavy-light orientation as scFv displayed ˜10-foldtighter than in the LH orientation binding to cells, consistent withELISA data. The nature of the mutations to eliminate the risk ofN106-based deamidation had a significant effect on T cell binding and oncytotoxicity. Altogether, the three different sets of back mutations,(defined by CD3B2030, CD3B2051, and CD3B2089) combined with mutation ofN106Q/A/G/S resulted in a panel of Cris-7 variants with EC50 for T cellbinding ranging from 3 to ˜300 nM, and correlated with EC50 forcytotoxicity from 0.012 to 3.5 nM. For T cell-redirecting bsAbs (bsTCE),weaker affinity T cell redirection, relative to the affinity for tumortargeting can allow design of the antibody to maximize efficacy whileminimizing toxicity associated with aberrant T cell activation,accumulation in secondary lymphoid organs, and cytokine-release-relatedtoxicity. Additionally Cris-7-derived scFvs in the “LH” orientation hadweaker binding to T cells, compared to HL orientation. Thus, this panelwas considered advantageous—and thus we selected a subset of the Cris-7variants to display a range of binding affinity to use in lead bsTCE.

TABLE 29 T cell binding and cytotoxicity analysis of Cris-7 × BCMA bsAbsusing H929 cells. H929 Binding, Killing T cell EC50, EC50, activation,Cris7 Variant nM pM % CD25 BC3B102.001 B2030 HL 3 NA NA BC3B106.001B2051 HL 6.8 NA NA BC3B111.001 B2089 HL 6.4 NA NA BC3B116.001 B2030 LH21.8 NA NA BC3B120.001 B2051 LH 100.4 NA NA BC3B125.001 B2089 LH 36.9 NANA BC3B51.001 B2030 HL NtoQ 1.9 12.9 23.4953116 BC3B109.001 B2051 HLNtoQ 13.8 42.6 110.55834 BC3B114.001 B2089 HL NtoQ 3.2 12.6 24.7547018BC3B53.001 B2030 LH NtoQ 19.6 27.8 56.6770832 BC3B123.001 B2051 LH NtoQ65 69.0 129.712199 BC3B128.001 B2089 LH NtoQ 42.9 35.4 81.8260112BC3B103.001 B2030 HL NtoA 4.6 92.1 153.584034 BC3B107.001 B2051 HL NtoA8.6 41.5 90.2349376 BC3B112.001 B2089 HL NtoA 300 NA NA BC3B117.001B2030 LH NtoA 60.9 148.6 279.030715 BC3B121.001 B2051 LH NtoA 73.3 100.2192.322866 BC3B126.001 B2089 LH NtoA 177.3 182.0 339.169276 BC3B104.001B2030 HL NtoG 64.2 236.6 448.804965 BC3B108.001 B2051 HL NtoG 11 85.3192.696137 BC3B113.001 B2089 HL NtoG 11.1 44.3 94.2622022 BC3B118.001B2030 LH NtoG 256.7 745.1 870.792549 BC3B122.001 B2051 LH NtoG 3004360.5 6136.07165 BC3B127.001 B2089 LH NtoG 300 805.9 1063.42224BC3B105.001 B2030 HL NtoS 20.7 172.2 272.296713 BC3B110.001 B2051 HLNtoS 31.2 780.5 1662.96965 BC3B115.001 B2089 HL NtoS 24 155.2 323.251509BC3B119.001 B2030 LH NtoS 300 10000.0 9999.99994 BC3B124.001 B2051 LHNtoS 300 NA NA BC3B129.001 B2089 LH NtoS 300 10000.0 9999.99994

TABLE 30 Functional activity of the bi-specific proteins. EC₅₀,Cytotoxicity Name BsAb Description (M) Cris7b GCDB131 HC1 (ZWA):N-Terminal 3.3364E−11 Cris7b-Fab; HC2 (ZWB): N- Terminal BCMB519-LH-scFvCris7- BC3B51 HC1 (ZWA): CD3B2030NtoQ 2.7847E−11 CD3B2030- HL scFv; HC2(ZWB): VH-N1060 BCMB519 Fab Cris7- BC3B53 HC1 (ZWA): CD3B2030NtoQ2.7847E−11 CD3B2030- LH scFv; HC2 (ZWB): VH-N1060 BCMB519 Fab Cris7-BC3B103 HC1 (ZWA): CD3B2030NtoA 9.213E−11 CD3B2030- HL scFv; HC2 (ZWB):VH-N106A BCMB519 Fab Cris7- BC3B117 HC1 (ZWA): CD3B2030NtoA 1.4858E−10CD3B2030- LH scFv; HC2 (ZWB): VH-N106A BCMB519 Fab Cris7- BC3B104 HC1(ZWA): CD3B2030NtoG 2.3659E−10 CD3B2030- HL scFv; HC2 (ZWB): VH-N106GBCMB519 Fab Cris7- BC3B118 HC1 (ZWA): CD3B2030NtoG 7.4505E−10 CD3B2030-LH scFv; HC2 (ZWB): VH-N106G BCMB519 Fab Cris7- BC3B105 HC1 (ZWA):CD3B2030NtoS 1.7224E−10 CD3B2030- HL scFv; HC2 (ZWB): VH-N106S BCMB519Fab Cris7- BC3B119 HC1 (ZWA): CD3B2030NtoS 1E−08 CD3B2030- LH scFv; HC2(ZWB): VH-N106S BCMB519 Fab Cris7- BC3B109 HC1 (ZWA): CD3B2051NtoQ4.2561E−11 CD3B2051- HL scFv; HC2 (ZWB): VH-N1060 BCMB519 Fab Cris7-BC3B123 HC1 (ZWA): CD3B2051NtoQ 6.9003E−11 CD3B2051- LH scFv; HC2 (ZWB):VH-N1060 BCMB519 Fab Cris7- BC3B107 HC1 (ZWA): CD3B2051NtoA 4.1534E−11CD3B2051- HL scFv; HC2 (ZWB): VH-N106A BCMB519 Fab Cris7- BC3B121 HC1(ZWA): CD3B2051NtoA 1.0015E−10 CD3B2051- LH scFv; HC2 (ZWB): VH-N106ABCMB519 Fab Cris7- BC3B108 HC1 (ZWA): CD3B2051NtoG 8.5329E−11 CD3B2051-HL scFv; HC2 (ZWB): VH-N106G BCMB519 Fab Cris7- BC3B122 HC1 (ZWA):CD3B2051NtoG 4.3605E−09 CD3B2051- LH scFv; HC2 (ZWB): VH-N106G BCMB519Fab Cris7- BC3B110 HC1 (ZWA): CD3B2051NtoS 7.8052E−10 CD3B2051- HL scFv;HC2 (ZWB): VH-N106S BCMB519 Fab Cris7- BC3B114 HC1 (ZWA): CD3B2089NtoQ1.255E−11 CD3B2089- HL scFv; HC2 (ZWB): VH-N1060 BCMB519 Fab Cris7-BC3B128 HC1 (ZWA): CD3B2089NtoQ 3.5408E−11 CD3B2089- LH scFv; HC2 (ZWB):VH-N1060 BCMB519 Fab Cris7- BC3B126 HC1 (ZWA): CD3B2089NtoA 1.8201E−10CD3B2089- LH scFv; HC2 (ZWB): VH-N106A BCMB519 Fab Cris7- BC3B113 HC1(ZWA): CD3B2089NtoG 4.4345E−11 CD3B2089- HL scFv; HC2 (ZWB): VH-N106GBCMB519 Fab Cris7- BC3B127 HC1 (ZWA): CD3B2089NtoG 8.059E−10 CD3B2089-LH scFv; HC2 (ZWB): VH-N106G BCMB519 Fab Cris7- BC3B115 HC1 (ZWA):CD3B2089NtoS 1.5516E−10 CD3B2089- HL scFv; HC2 (ZWB): VH-N106S BCMB519Fab Cris7- BC3B129 HC1 (ZWA): CD3B2089NtoS N.D. CD3B2089- LH scFv; HC2(ZWB): VH-N106S BCMB519 Fab

Example 3: Expression and Purification of Bispecific CD79b×CD3 andTrispecific CD79b×CD20×CD3 Antibodies

The CD79b×CD3 bispecific antibody (bsAb) is an immunoglobulin (Ig) G1bispecific antibody that can bind simultaneously or individually to thecluster of differentiation (CD) 3 receptor complex on T lymphocytes andto CD79b on B lymphocytes. The CD79b×CD20×CD3 trispecific antibody is animmunoglobulin (Ig) G1 trispecific antibody that can bind simultaneouslyor individually to the CD3 receptor complex on T lymphocytes, and to theCD20 receptor complex on B lymphocytes and to the CD79b receptor complexon B lymphocytes. The antibody has mutations which reduce Fc binding toa Fcγ receptor and heterodimerization has been enhanced using theknobs-in-holes platform mutations. The trispecific antibody wasdeveloped to evaluate the therapeutic potential of dual targeting CD20and CD79b for T cell redirection. An illustration of an exemplaryCD79b×CD20×CD3 antibody is depicted in FIG. 6 .

Table 31 provides a summary of examples of some CD79b×CD20×CD3trispecific antibodies described herein:

TABLE 31 Exemplary CD79b × CD20 × CD3 Trispecific antibodies HC1 LC HC2Amino HC1 Amino Amino HC2 acid DNA acid LC DNA acid DNA HC1/LC sequencesequence sequence sequence sequence sequence (CD79b SEQ ID SEQ ID SEQ IDSEQ ID HC2 (CD3- SEQ ID SEQ ID ID arm) NO NO NO NO CD20 arm) NO NOC923B38 CD9B374 1489 1490 1491 1492 CD3B2030- 1463 1464 N106A- scFv-LH-C20B22 C923B74 CD9B330- 1493 1494 1495 1496 CD3B2030- 1463 1464 N31SN106A- scFv-LH- C20B22 C923B99 CD9B643 1497 1498 1499 1500 CD3B2030-1463 1464 N106A- scFv-LH- C20B22 C923B36 CD9B374 1489 1490 1491 1492CD3B2089- 1465 1466 N106G- scFv-LH- C20B22 C923B73 CD9B330- 1493 14941495 1496 CD3B2089- 1465 1466 N31S N106G- scFv-LH- C20B22 C923B95CD9B643 1497 1498 1499 1500 CD3B2089- 1465 1466 N106G- scFv-LH- C20B22C923B138 CD9B643 1497 1498 1499 1501 CD3W245- 1467 1468 scFv LH- C20B22C923B139 CD9B643 1497 1498 1499 1501 CD3B2089- 1469 1470 N106G-scFvHL-C20B22 C923B140 CD9B643 1497 1498 1499 1501 CD3W245- 1471 1472 scFvLH- 5O10GL C923B141 CD9B643 1497 1498 1499 1501 CD3W245- 1473 1474 scFvLH- 4A16GL C923B142 CD9B643 1497 1498 1499 1501 CD3B2030- 1475 1476N106A-LH scFv- 5O10GL C923B143 CD9B643 1497 1498 1499 1501 CD3B2030-1477 1478 N106A-LH scFv- 4A16GL C923B144 CD9B643 1497 1498 1499 1501CD3B2089- 1479 1480 N106G-HL scFv- 5O10GL C923B145 CD9B643 1497 14981499 1501 CD3B2089- 1481 1482 N106G-HL scFv- 4A16GL C923B147 CD9B6431502 1503 1499 1500 CD3B2030- 1483 1484 N106A-LH scFv- 4A16GL C923B168CD9B374 1489 1490 1491 1492 CD3W245- 1485 1486 scFv LH- C20B648 LHC923B169 CD9B374 1489 1490 1491 1492 CD3B2030- 1487 1488 N106A-LH-C20B648 LH

trispecific Ab CD3-CD20 arm SEQ ID NO: 1463EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGSEGKSSGSGSESKSTGGSAIQLTQSPSSLSASVGDRVTITCRASSSVSYIHWFQQKPGKAPKPLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQWTSNPPTFGQGTKLEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGAIYPGNGDTSYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYY CARSTYYGGDWYFNVWGQGTLVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1464GAGATCGTGCTGACCCAGTCTCCTGCCACACTGAGTGCTTCTCCAGGCGAGAGAGTGACCCTGTCCTGCTCCGCTTCCTCCTCCGTGTCCTACATGAACTGGTATCAGCAGAAGCCCGGCCAGGCTCCTCGGAGATGGATCTACGACTCTTCCAAGCTGGCCTCTGGTGTGCCAGCCAGATTTTCTGGCTCTGGCTCCGGCAGAGACTATACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTACTGCCAGCAGTGGTCTAGGAACCCTCCTACCTTTGGCGGAGGCACCAAGGTGGAAATCAAGGGCGGATCTGAGGGAAAGTCCAGCGGCTCCGGCAGCGAAAGCAAGTCCACCGGCGGAAGCCAGGTTCAACTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCTTCCGGCTACACCTTTACCAGATCCACCATGCACTGGGTCAAGCAGGCCCCTGGACAAGGCTTGGAGTGGATCGGCTACATCAACCCCAGCTCCGCCTACACCAACTACAACCAGAAATTCCAGGGCAGAGTGACCCTGACCGCCGACAAGTCTACCTCCACCGCCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTGCGCCTCTCCTCAGGTCCACTACGACTACGCCGGCTTTCCTTATTGGGGCCAGGGCACACTGGTCACCGTTTCTTCTGAGCCCAAATCTAGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGTCTCTCTCCCTGTCTCCGGGAAAAGGAGGGAGCGAGGGAAAGTCCAGCGGAAGCGGCTCTGAGTCCAAATCCACCGGAGGGAGCGCCATTCAGCTGACCCAGTCTCCATCCTCTCTGTCCGCCTCTGTGGGCGACAGAGTGACAATTACCTGCCGGGCCTCCTCCTCCGTGTCCTACATCCATTGGTTCCAGCAGAAGCCCGGCAAGGCCCCTAAGCCTCTGATCTACGCCACCTCCAATCTGGCCTCTGGCGTGCCCTCCAGATTTTCCGGATCTGGCTCTGGAACCGACTTTACCCTGACAATCTCCAGCCTGCAGCCTGAGGACTTCGCCACCTACTACTGTCAGCAGTGGACCAGCAATCCTCCTACCTTTGGCCAGGGCACCAAGCTGGAAATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAGGTTCAGCTGGTTCAGTCTGGTGCCGAAGTGAAGAAACCTGGCTCCTCCGTGAAAGTGTCCTGCAAGGCTTCCGGCTACACTTTTACCAGCTACAACATGCACTGGGTCCGACAGGCCCCTGGACAAGGATTGGAATGGATGGGCGCTATCTACCCCGGCAACGGCGATACCTCTTACGCCCAGAAATTCCAGGGCAGAGTGACCATCACCGCCGACAAGTCTACCTCCACCGCCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTGCGCCCGGTCTACCTATTATGGCGGCGACTGGTACTTCAACGTGTGGGGCCAGGGAACCCTGGTCACAGTCTCTTCT trispecific Ab CD3-CD20 armSEQ ID NO: 1465 EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGSEGKSSGSGSESKSTGGSAIQLTQSPSSLSASVGDRVTITCRASSSVSYIHWFQQKPGKAPKPLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQWTSNPPTFGQGTKLEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGAIYPGNGDTSYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYY CARSTYYGGDWYFNVWGQGTLVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1466GAGATCGTGCTGACCCAGTCTCCTGCCACACTGAGTGCTTCTCCAGGCGAGAGAGTGACCCTGTCCTGCTCCGCTTCCTCCTCCGTGTCCTACATGAACTGGTATCAGCAGAAGCCCGGCCAGGCTCCTCGGAGATGGATCTACGACTCTTCCAAGCTGGCCTCTGGTGTGCCAGCCAGATTTTCTGGCTCTGGCTCCGGCAGAGACTATACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTACTGCCAGCAGTGGTCTAGGAACCCTCCTACCTTTGGCGGAGGCACCAAGGTGGAAATCAAGGGCGGATCTGAGGGAAAGTCCAGCGGCTCCGGCAGCGAAAGCAAGTCCACCGGCGGAAGCCAGGTTCAACTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCTCCTCCGTGAAAGTGTCCTGCAAGGCTTCCGGCTACACTTTTACCAGATCCACCATGCACTGGGTCCGACAGGCTCCAGGACAAGGCTTGGAGTGGATGGGCTACATCAACCCCAGCTCCGCCTACACCAACTACGCCCAGAAATTCCAGGGCAGAGTGACCCTGACCGCCGACAAGTCTACCTCCACCGCCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTGCGCTTCTCCTCAGGTGCACTACGACTACGGCGGCTTTCCTTATTGGGGCCAGGGCACACTGGTCACCGTTTCTTCTGAGCCCAAATCTAGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGTCTCTCTCCCTGTCTCCGGGAAAAGGAGGGAGCGAGGGAAAGTCCAGCGGAAGCGGCTCTGAGTCCAAATCCACCGGAGGGAGCGCCATTCAGCTGACCCAGTCTCCATCCTCTCTGTCCGCCTCTGTGGGCGACAGAGTGACAATTACCTGCCGGGCCTCCTCCTCCGTGTCCTACATCCATTGGTTCCAGCAGAAGCCCGGCAAGGCCCCTAAGCCTCTGATCTACGCCACCTCCAATCTGGCCTCTGGCGTGCCCTCCAGATTTTCCGGATCTGGCTCTGGAACCGACTTTACCCTGACAATCTCCAGCCTGCAGCCTGAGGACTTCGCCACCTACTACTGTCAGCAGTGGACCAGCAATCCTCCTACCTTTGGCCAGGGCACCAAGCTGGAAATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAGGTTCAGCTGGTTCAGTCTGGTGCCGAAGTGAAGAAACCTGGCTCCTCCGTGAAAGTGTCCTGCAAGGCTTCCGGCTACACTTTTACCAGCTACAACATGCACTGGGTCCGACAGGCCCCTGGACAAGGATTGGAATGGATGGGCGCTATCTACCCCGGCAACGGCGATACCTCTTACGCCCAGAAATTCCAGGGCAGAGTGACCATCACCGCCGACAAGTCTACCTCCACCGCCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTGCGCCCGGTCTACCTATTATGGCGGCGACTGGTACTTCAACGTGTGGGGCCAGGGAACCCTGGTCACAGTCTCTTCT trispecific Ab CD3-CD20 armSEQ ID NO: 1467 DIQMTQSPSSLSASVGDRVTITCRARQSIGTAIHWYQQKPGKAPKLLIKYASESISGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSGSWPYTFGQGTKLEIKGGSEGKSSGSGSESKSTGGSEVQLVESGGGLVKPGGSLRLSCAASGFTFSRYNMNWVRQAPGKGLEWVSSISTSSNYIYYADSVKGRFTFSRDNAKNSLDLQMSGLRAEDTAIYYCTRGWGPFDYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSAIQLTQSPSSLSASVGDRVTITCRASSSVSYIHWFQQKPGKAPKPLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQWTSNPPTFGQGTKLEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGAIYPGNGDTSYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSTYYGGDWYFN VWGQGTLVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1468GACATACAAATGACACAATCACCCTCTTCTCTTTCTGCAAGCGTTGGCGACCGTGTCACTATCACTTGTCGAGCCCGCCAGTCCATAGGTACTGCCATTCACTGGTATCAACAGAAGCCTGGCAAGGCTCCCAAACTCCTGATTAAGTATGCCAGCGAGAGCATTTCCGGCGTACCTTCAAGATTTTCCGGCTCCGGTAGTGGGACAGATTTCACTCTCACTATATCTAGCCTCCAACCAGAAGATTTCGCCACTTACTACTGTCAACAATCAGGTTCATGGCCTTACACTTTCGGCCAGGGGACAAAATTGGAGATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCGAGGTGCAACTGGTGGAGTCTGGGGGAGGCCTGGTCAAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGATATAACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCATCCATTAGTACTAGTAGTAATTACATATACTACGCAGACTCAGTGAAGGGCCGATTCACCTTCTCCAGAGACAACGCCAAGAACTCACTGGATCTGCAAATGAGCGGCCTGAGAGCCGAGGACACGGCTATTTATTACTGTACGAGAGGCTGGGGGCCTTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGAGCCCAAATCTAGCGACAAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAGGAGGCGGAGGGAGTGGCGGGGGAGGCTCTGCAATCCAACTAACTCAAAGTCCAAGTAGTCTGTCTGCTTCCGTGGGCGACAGAGTGACAATCACCTGTAGAGCCTCCAGCAGCGTCTCCTACATCCACTGGTTCCAGCAAAAACCTGGCAAGGCCCCTAAGCCTCTGATCTACGCCACCTCCAACCTGGCCTCTGGCGTGCCCTCTCGGTTCTCCGGCTCTGGCTCCGGAACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGATTTTGCTACCTACTACTGCCAGCAGTGGACCTCTAACCCTCCAACATTCGGCCAGGGCACCAAGCTGGAAATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAAGTGCAATTAGTGCAAAGTGGTGCAGAAGTCAAGAAGCCTGGAAGCTCCGTGAAAGTGTCCTGCAAGGCCTCTGGCTACACCTTTACCTCCTACAACATGCACTGGGTGCGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCGCTATCTACCCCGGCAACGGCGATACCTCTTACGCCCAGAAGTTCCAGGGCAGAGTGACCATCACCGCCGACAAGTCCACATCTACAGCCTACATGGAACTGTCCTCCCTGCGGTCCGAGGACACCGCTGTGTACTATTGTGCCAGATCTACCTACTACGGCGGCGACTGGTACTTCAACGTGTGGGGCCAAGGAACCCTGGTGACCGTGTCTAGC trispecific Ab CD3-CD20 armSEQ ID NO: 1469 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSAIQLTQSPSSLSASVGDRVTITCRASSSVSYIHWFQQKPGKAPKPLIYATSNLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQWTSNPPTFGQGTKLEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGAIYPGNGDTSYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSTYYGGD WYFNVWGQGTLVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1470CAGGTGCAGCTGGTGCAGAGCGGCGCCGAGGTGAAGAAGCCTGGCAGCAGCGTGAAGGTGAGCTGTAAGGCCAGCGGCTACACTTTCACTAGGAGCACTATGCACTGGGTGAGGCAGGCCCCTGGCCAGGGCCTGGAGTGGATGGGCTACATCAATCCTAGCAGCGCCTACACTAATTACGCCCAGAAGTTCCAGGGCAGGGTGACTCTGACTGCCGATAAGAGCACTAGCACTGCCTACATGGAGCTGAGCAGCCTGAGGAGCGAGGATACTGCCGTGTACTACTGTGCCAGCCCTCAGGTGCACTACGATTACGGCGGCTTCCCTTACTGGGGCCAGGGCACTCTGGTGACTGTGAGCAGCGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCGAGATCGTGCTGACTCAGAGCCCTGCCACTCTGAGCGCCAGCCCTGGCGAGAGGGTGACTCTGAGCTGTAGCGCCAGCAGCAGCGTGAGCTACATGAATTGGTACCAGCAGAAGCCTGGCCAGGCCCCTAGGAGGTGGATCTACGATAGCAGCAAGCTGGCCAGCGGCGTGCCTGCCAGGTTCAGCGGCAGCGGCAGCGGCAGGGATTACACTCTGACTATCAGCAGCCTGGAGCCTGAGGATTTCGCCGTGTACTACTGTCAGCAGTGGAGCAGGAATCCTCCTACTTTCGGCGGCGGCACTAAGGTGGAGATCAAGGAGCCCAAATCTAGCGACAAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAGGAGGCGGAGGGAGTGGCGGGGGAGGCTCTGCAATCCAACTAACTCAAAGTCCAAGTAGTCTGTCTGCTTCCGTGGGCGACAGAGTGACAATCACCTGTAGAGCCTCCAGCAGCGTCTCCTACATCCACTGGTTCCAGCAAAAACCTGGCAAGGCCCCTAAGCCTCTGATCTACGCCACCTCCAACCTGGCCTCTGGCGTGCCCTCTCGGTTCTCCGGCTCTGGCTCCGGAACCGACTTCACCCTGACCATCTCCAGCCTGCAGCCTGAGGATTTTGCTACCTACTACTGCCAGCAGTGGACCTCTAACCCTCCAACATTCGGCCAGGGCACCAAGCTGGAAATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAAGTGCAATTAGTGCAAAGTGGTGCAGAAGTCAAGAAGCCTGGAAGCTCCGTGAAAGTGTCCTGCAAGGCCTCTGGCTACACCTTTACCTCCTACAACATGCACTGGGTGCGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCGCTATCTACCCCGGCAACGGCGATACCTCTTACGCCCAGAAGTTCCAGGGCAGAGTGACCATCACCGCCGACAAGTCCACATCTACAGCCTACATGGAACTGTCCTCCCTGCGGTCCGAGGACACCGCTGTGTACTATTGTGCCAGATCTACCTACTACGGCGGCGACTGGTACTTCAACGTGTGGGGCCAAGGAACCCTGGTGACCGTGTCT AGCtrispecific Ab CD3-CD20 arm SEQ ID NO: 1471DIQMTQSPSSLSASVGDRVTITCRARQSIGTAIHWYQQKPGKAPKLLIKYASESISGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSGSWPYTFGQGTKLEIKGGSEGKSSGSGSESKSTGGSEVQLVESGGGLVKPGGSLRLSCAASGFTFSRYNMNWVRQAPGKGLEWVSSISTSSNYIYYADSVKGRFTFSRDNAKNSLDLQMSGLRAEDTAIYYCTRGWGPFDYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASSSVSYMHWYQQKPGSSPQVWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDTATYYCQQWIFNPPTFGSGTKLEIRGGSEGKSSGSGSESKSTGGSQAYLQQSGAELVRPGASVKMSCKASGYTFTSYNMHWVKQTPRQGLEWIGAIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCARVYYGSNYWYFD VWGTGTTVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1472GACATACAAATGACACAATCACCCTCTTCTCTTTCTGCAAGCGTTGGCGACCGTGTCACTATCACTTGTCGAGCCCGCCAGTCCATAGGTACTGCCATTCACTGGTATCAACAGAAGCCTGGCAAGGCTCCCAAACTCCTGATTAAGTATGCCAGCGAGAGCATTTCCGGCGTACCTTCAAGATTTTCCGGCTCCGGTAGTGGGACAGATTTCACTCTCACTATATCTAGCCTCCAACCAGAAGATTTCGCCACTTACTACTGTCAACAATCAGGTTCATGGCCTTACACTTTCGGCCAGGGGACAAAATTGGAGATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCGAGGTGCAACTGGTGGAGTCTGGGGGAGGCCTGGTCAAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGATATAACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCATCCATTAGTACTAGTAGTAATTACATATACTACGCAGACTCAGTGAAGGGCCGATTCACCTTCTCCAGAGACAACGCCAAGAACTCACTGGATCTGCAAATGAGCGGCCTGAGAGCCGAGGACACGGCTATTTATTACTGTACGAGAGGCTGGGGGCCTTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGAGCCCAAATCTAGCGACAAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAGGAGGCGGAGGGAGTGGCGGGGGAGGCTCTCAAATAGTCCTTTCACAGTCCCCAGCTATTCTTTCAGCCTCTCCCGGTGAAAAGGTTACAATGACCTGCCGGGCAAGCTCCAGTGTCTCATATATGCACTGGTACCAACAAAAACCTGGCAGTAGTCCTCAGGTGTGGATCTACGCTACAAGCAATCTCGCTTCCGGGGTTCCCGTGAGGTTTAGCGGAAGCGGGTCTGGAACTAGTTATTCCTTGACAATTAGTCGGGTTGAAGCCGAGGACACCGCCACTTACTATTGCCAACAGTGGATATTCAATCCACCCACCTTCGGTTCAGGTACCAAGCTCGAAATCCGTGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAAGCATATCTGCAACAGAGCGGAGCTGAGCTGGTTCGGCCTGGCGCCTCTGTAAAAATGAGTTGCAAGGCCAGTGGTTATACATTCACATCATATAATATGCACTGGGTAAAGCAAACTCCCCGACAGGGGCTTGAATGGATTGGCGCAATCTATCCCGGCAATGGGGATACATCCTACAATCAGAAATTCAAGGGCAAGGCAACACTGACCGTTGACAAATCCTCATCAACAGCCTACATGCAGCTCAGTTCCCTCACTAGCGAAGATTCTGCTGTGTATTTCTGTGCAAGGGTGTATTATGGTTCTAATTACTGGTATTTCGATGTTTGGGGAACCGGAACTACCGTAACTGTTTCTAGC trispecific Ab CD3-CD20 armSEQ ID NO: 1473 DIQMTQSPSSLSASVGDRVTITCRARQSIGTAIHWYQQKPGKAPKLLIKYASESISGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSGSWPYTFGQGTKLEIKGGSEGKSSGSGSESKSTGGSEVQLVESGGGLVKPGGSLRLSCAASGFTFSRYNMNWVRQAPGKGLEWVSSISTSSNYIYYADSVKGRFTFSRDNAKNSLDLQMSGLRAEDTAIYYCTRGWGPFDYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASLSVSSMHWYQQKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWIFNPPTFGGGTKLEIKGGSEGKSSGSGSESKSTGGSQAYLQQSGAELVRPGASVKMSCKTSGYTFSSYNMHWVKQTPRQALEWIGAIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCTRSNYYGSSGWYF DVWGTGTTVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1474GACATACAAATGACACAATCACCCTCTTCTCTTTCTGCAAGCGTTGGCGACCGTGTCACTATCACTTGTCGAGCCCGCCAGTCCATAGGTACTGCCATTCACTGGTATCAACAGAAGCCTGGCAAGGCTCCCAAACTCCTGATTAAGTATGCCAGCGAGAGCATTTCCGGCGTACCTTCAAGATTTTCCGGCTCCGGTAGTGGGACAGATTTCACTCTCACTATATCTAGCCTCCAACCAGAAGATTTCGCCACTTACTACTGTCAACAATCAGGTTCATGGCCTTACACTTTCGGCCAGGGGACAAAATTGGAGATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCGAGGTGCAACTGGTGGAGTCTGGGGGAGGCCTGGTCAAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGATATAACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCATCCATTAGTACTAGTAGTAATTACATATACTACGCAGACTCAGTGAAGGGCCGATTCACCTTCTCCAGAGACAACGCCAAGAACTCACTGGATCTGCAAATGAGCGGCCTGAGAGCCGAGGACACGGCTATTTATTACTGTACGAGAGGCTGGGGGCCTTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGAGCCCAAATCTAGCGACAAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAGGAGGCGGAGGGAGTGGCGGGGGAGGCTCTCAGATTGTCCTGAGCCAATCCCCAGCAATTCTGAGTGCTAGCCCTGGAGAGAAGGTAACAATGACTTGTCGGGCATCCCTTAGCGTCTCATCCATGCATTGGTATCAACAAAAGCCAGGTTCATCTCCAAAACCCTGGATTTACGCTACATCTAACCTGGCATCTGGGGTGCCTGCCAGATTTAGTGGATCTGGTTCCGGCACATCATATTCCCTTACAATCAGCCGAGTGGAAGCCGAGGATGCTGCAACCTATTACTGTCAACAATGGATATTTAACCCTCCCACCTTTGGGGGTGGGACTAAACTCGAAATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAAGCCTATCTTCAACAATCTGGGGCTGAGCTTGTCCGGCCAGGAGCCTCCGTCAAAATGAGCTGCAAAACCTCAGGTTATACTTTTAGTAGCTATAACATGCATTGGGTAAAACAAACCCCCCGACAAGCATTGGAGTGGATAGGGGCCATATACCCCGGCAATGGAGACACAAGTTACAACCAGAAGTTTAAAGGCAAAGCTACACTCACAGTTGACAAATCCTCAAGTACTGCTTATATGCAACTCTCCTCTCTCACTTCCGAAGACAGTGCCGTATATTTTTGCACTCGGTCCAATTACTATGGATCTAGTGGCTGGTACTTTGACGTTTGGGGCACTGGGACAACTGTTACAGTGTCCAGC trispecific Ab CD3-CD20 armSEQ ID NO: 1475 EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASSSVSYMHWYQQKPGSSPQVWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDTATYYCQQWIFNPPTFGSGTKLEIRGGSEGKSSGSGSESKSTGGSQAYLQQSGAELVRPGASVKMSCKASGYTFTSYNMHWVKQTPRQGLEWIGAIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCARVYYGSNY WYFDVWGTGTTVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1476GAGATCGTGCTGACTCAGAGCCCTGCCACTCTGAGCGCCAGCCCTGGCGAGAGGGTGACTCTGAGCTGTAGCGCCAGCAGCAGCGTGAGCTACATGAATTGGTACCAGCAGAAGCCTGGCCAGGCCCCTAGGAGGTGGATCTACGATAGCAGCAAGCTGGCCAGCGGCGTGCCTGCCAGGTTCAGCGGCAGCGGCAGCGGCAGGGATTACACTCTGACTATCAGCAGCCTGGAGCCTGAGGATTTCGCCGTGTACTACTGTCAGCAGTGGAGCAGGAATCCTCCTACTTTCGGCGGCGGCACTAAGGTGGAGATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAGGTGCAGCTGGTGCAGAGCGGCGCCGAGGTGAAGAAGCCTGGCAGCAGCGTGAAGGTGAGCTGTAAGGCCAGCGGCTACACTTTCACTAGGAGCACTATGCACTGGGTGAAGCAGGCCCCTGGCCAGGGCCTGGAGTGGATCGGCTACATCAATCCTAGCAGCGCCTACACTAATTACAATCAGAAGTTCCAGGGCAGGGTGACTCTGACTGCCGATAAGAGCACTAGCACTGCCTACATGGAGCTGAGCAGCCTGAGGAGCGAGGATACTGCCGTGTACTACTGTGCCAGCCCTCAGGTGCACTACGATTACGCCGGCTTCCCTTACTGGGGCCAGGGCACTCTGGTGACTGTGAGCAGCGAGCCCAAATCTAGCGACAAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAGGAGGCGGAGGGAGTGGCGGGGGAGGCTCTCAAATAGTCCTTTCACAGTCCCCAGCTATTCTTTCAGCCTCTCCCGGTGAAAAGGTTACAATGACCTGCCGGGCAAGCTCCAGTGTCTCATATATGCACTGGTACCAACAAAAACCTGGCAGTAGTCCTCAGGTGTGGATCTACGCTACAAGCAATCTCGCTTCCGGGGTTCCCGTGAGGTTTAGCGGAAGCGGGTCTGGAACTAGTTATTCCTTGACAATTAGTCGGGTTGAAGCCGAGGACACCGCCACTTACTATTGCCAACAGTGGATATTCAATCCACCCACCTTCGGTTCAGGTACCAAGCTCGAAATCCGTGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAAGCATATCTGCAACAGAGCGGAGCTGAGCTGGTTCGGCCTGGCGCCTCTGTAAAAATGAGTTGCAAGGCCAGTGGTTATACATTCACATCATATAATATGCACTGGGTAAAGCAAACTCCCCGACAGGGGCTTGAATGGATTGGCGCAATCTATCCCGGCAATGGGGATACATCCTACAATCAGAAATTCAAGGGCAAGGCAACACTGACCGTTGACAAATCCTCATCAACAGCCTACATGCAGCTCAGTTCCCTCACTAGCGAAGATTCTGCTGTGTATTTCTGTGCAAGGGTGTATTATGGTTCTAATTACTGGTATTTCGATGTTTGGGGAACCGGAACTACCGTAACTGTTTCT AGCtrispecific Ab CD3-CD20 arm SEQ ID NO: 1477EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASLSVSSMHWYQQKPGSSPKPWIYATSNLASGVPARESGSGSGTSYSLTISRVEAEDAATYYCQQWIFNPPTFGGGTKLEIKGGSEGKSSGSGSESKSTGGSQAYLQQSGAELVRPGASVKMSCKTSGYTFSSYNMHWVKQTPRQALEWIGAIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCTRSNYYGSS GWYFDVWGTGTTVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1478GAGATCGTGCTGACTCAGAGCCCTGCCACTCTGAGCGCCAGCCCTGGCGAGAGGGTGACTCTGAGCTGTAGCGCCAGCAGCAGCGTGAGCTACATGAATTGGTACCAGCAGAAGCCTGGCCAGGCCCCTAGGAGGTGGATCTACGATAGCAGCAAGCTGGCCAGCGGCGTGCCTGCCAGGTTCAGCGGCAGCGGCAGCGGCAGGGATTACACTCTGACTATCAGCAGCCTGGAGCCTGAGGATTTCGCCGTGTACTACTGTCAGCAGTGGAGCAGGAATCCTCCTACTTTCGGCGGCGGCACTAAGGTGGAGATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAGGTGCAGCTGGTGCAGAGCGGCGCCGAGGTGAAGAAGCCTGGCAGCAGCGTGAAGGTGAGCTGTAAGGCCAGCGGCTACACTTTCACTAGGAGCACTATGCACTGGGTGAAGCAGGCCCCTGGCCAGGGCCTGGAGTGGATCGGCTACATCAATCCTAGCAGCGCCTACACTAATTACAATCAGAAGTTCCAGGGCAGGGTGACTCTGACTGCCGATAAGAGCACTAGCACTGCCTACATGGAGCTGAGCAGCCTGAGGAGCGAGGATACTGCCGTGTACTACTGTGCCAGCCCTCAGGTGCACTACGATTACGCCGGCTTCCCTTACTGGGGCCAGGGCACTCTGGTGACTGTGAGCAGCGAGCCCAAATCTAGCGACAAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAGGAGGCGGAGGGAGTGGCGGGGGAGGCTCTCAGATTGTCCTGAGCCAATCCCCAGCAATTCTGAGTGCTAGCCCTGGAGAGAAGGTAACAATGACTTGTCGGGCATCCCTTAGCGTCTCATCCATGCATTGGTATCAACAAAAGCCAGGTTCATCTCCAAAACCCTGGATTTACGCTACATCTAACCTGGCATCTGGGGTGCCTGCCAGATTTAGTGGATCTGGTTCCGGCACATCATATTCCCTTACAATCAGCCGAGTGGAAGCCGAGGATGCTGCAACCTATTACTGTCAACAATGGATATTTAACCCTCCCACCTTTGGGGGTGGGACTAAACTCGAAATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAAGCCTATCTTCAACAATCTGGGGCTGAGCTTGTCCGGCCAGGAGCCTCCGTCAAAATGAGCTGCAAAACCTCAGGTTATACTTTTAGTAGCTATAACATGCATTGGGTAAAACAAACCCCCCGACAAGCATTGGAGTGGATAGGGGCCATATACCCCGGCAATGGAGACACAAGTTACAACCAGAAGTTTAAAGGCAAAGCTACACTCACAGTTGACAAATCCTCAAGTACTGCTTATATGCAACTCTCCTCTCTCACTTCCGAAGACAGTGCCGTATATTTTTGCACTCGGTCCAATTACTATGGATCTAGTGGCTGGTACTTTGACGTTTGGGGCACTGGGACAACTGTTACAGTG TCCAGCtrispecific Ab CD3-CD20 arm SEQ ID NO: 1479QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASSSVSYMHWYQQKPGSSPQVWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDTATYYCQQWIFNPPTFGSGTKLEIRGGSEGKSSGSGSESKSTGGSQAYLQQSGAELVRPGASVKMSCKASGYTFTSYNMHWVKQTPRQGLEWIGAIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCARVYYGSNY WYFDVWGTGTTVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1480CAGGTGCAGCTGGTGCAGAGCGGCGCCGAGGTGAAGAAGCCTGGCAGCAGCGTGAAGGTGAGCTGTAAGGCCAGCGGCTACACTTTCACTAGGAGCACTATGCACTGGGTGAGGCAGGCCCCTGGCCAGGGCCTGGAGTGGATGGGCTACATCAATCCTAGCAGCGCCTACACTAATTACGCCCAGAAGTTCCAGGGCAGGGTGACTCTGACTGCCGATAAGAGCACTAGCACTGCCTACATGGAGCTGAGCAGCCTGAGGAGCGAGGATACTGCCGTGTACTACTGTGCCAGCCCTCAGGTGCACTACGATTACGGCGGCTTCCCTTACTGGGGCCAGGGCACTCTGGTGACTGTGAGCAGCGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCGAGATCGTGCTGACTCAGAGCCCTGCCACTCTGAGCGCCAGCCCTGGCGAGAGGGTGACTCTGAGCTGTAGCGCCAGCAGCAGCGTGAGCTACATGAATTGGTACCAGCAGAAGCCTGGCCAGGCCCCTAGGAGGTGGATCTACGATAGCAGCAAGCTGGCCAGCGGCGTGCCTGCCAGGTTCAGCGGCAGCGGCAGCGGCAGGGATTACACTCTGACTATCAGCAGCCTGGAGCCTGAGGATTTCGCCGTGTACTACTGTCAGCAGTGGAGCAGGAATCCTCCTACTTTCGGCGGCGGCACTAAGGTGGAGATCAAGGAGCCCAAATCTAGCGACAAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAGGAGGCGGAGGGAGTGGCGGGGGAGGCTCTCAAATAGTCCTTTCACAGTCCCCAGCTATTCTTTCAGCCTCTCCCGGTGAAAAGGTTACAATGACCTGCCGGGCAAGCTCCAGTGTCTCATATATGCACTGGTACCAACAAAAACCTGGCAGTAGTCCTCAGGTGTGGATCTACGCTACAAGCAATCTCGCTTCCGGGGTTCCCGTGAGGTTTAGCGGAAGCGGGTCTGGAACTAGTTATTCCTTGACAATTAGTCGGGTTGAAGCCGAGGACACCGCCACTTACTATTGCCAACAGTGGATATTCAATCCACCCACCTTCGGTTCAGGTACCAAGCTCGAAATCCGTGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAAGCATATCTGCAACAGAGCGGAGCTGAGCTGGTTCGGCCTGGCGCCTCTGTAAAAATGAGTTGCAAGGCCAGTGGTTATACATTCACATCATATAATATGCACTGGGTAAAGCAAACTCCCCGACAGGGGCTTGAATGGATTGGCGCAATCTATCCCGGCAATGGGGATACATCCTACAATCAGAAATTCAAGGGCAAGGCAACACTGACCGTTGACAAATCCTCATCAACAGCCTACATGCAGCTCAGTTCCCTCACTAGCGAAGATTCTGCTGTGTATTTCTGTGCAAGGGTGTATTATGGTTCTAATTACTGGTATTTCGATGTTTGGGGAACCGGAACTACCGTAACTGTTTCT AGCtrispecific Ab CD3-CD20 arm SEQ ID NO: 1481QVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSGGSEGKSSGSGSESKSTGGSEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASLSVSSMHWYQQKPGSSPKPWIYATSNLASGVPARESGSGSGTSYSLTISRVEAEDAATYYCQQWIFNPPTFGGGTKLEIKGGSEGKSSGSGSESKSTGGSQAYLQQSGAELVRPGASVKMSCKTSGYTFSSYNMHWVKQTPRQALEWIGAIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCTRSNYYGSS GWYFDVWGTGTTVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1482CAGGTGCAGCTGGTGCAGAGCGGCGCCGAGGTGAAGAAGCCTGGCAGCAGCGTGAAGGTGAGCTGTAAGGCCAGCGGCTACACTTTCACTAGGAGCACTATGCACTGGGTGAGGCAGGCCCCTGGCCAGGGCCTGGAGTGGATGGGCTACATCAATCCTAGCAGCGCCTACACTAATTACGCCCAGAAGTTCCAGGGCAGGGTGACTCTGACTGCCGATAAGAGCACTAGCACTGCCTACATGGAGCTGAGCAGCCTGAGGAGCGAGGATACTGCCGTGTACTACTGTGCCAGCCCTCAGGTGCACTACGATTACGGCGGCTTCCCTTACTGGGGCCAGGGCACTCTGGTGACTGTGAGCAGCGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCGAGATCGTGCTGACTCAGAGCCCTGCCACTCTGAGCGCCAGCCCTGGCGAGAGGGTGACTCTGAGCTGTAGCGCCAGCAGCAGCGTGAGCTACATGAATTGGTACCAGCAGAAGCCTGGCCAGGCCCCTAGGAGGTGGATCTACGATAGCAGCAAGCTGGCCAGCGGCGTGCCTGCCAGGTTCAGCGGCAGCGGCAGCGGCAGGGATTACACTCTGACTATCAGCAGCCTGGAGCCTGAGGATTTCGCCGTGTACTACTGTCAGCAGTGGAGCAGGAATCCTCCTACTTTCGGCGGCGGCACTAAGGTGGAGATCAAGGAGCCCAAATCTAGCGACAAAACTCACACATGTCCACCGTGCCCAGCACCTGAAGCAGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGTCTAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAAGGAGGCGGAGGGAGTGGCGGGGGAGGCTCTCAGATTGTCCTGAGCCAATCCCCAGCAATTCTGAGTGCTAGCCCTGGAGAGAAGGTAACAATGACTTGTCGGGCATCCCTTAGCGTCTCATCCATGCATTGGTATCAACAAAAGCCAGGTTCATCTCCAAAACCCTGGATTTACGCTACATCTAACCTGGCATCTGGGGTGCCTGCCAGATTTAGTGGATCTGGTTCCGGCACATCATATTCCCTTACAATCAGCCGAGTGGAAGCCGAGGATGCTGCAACCTATTACTGTCAACAATGGATATTTAACCCTCCCACCTTTGGGGGTGGGACTAAACTCGAAATCAAGGGCGGCTCCGAGGGCAAGAGCAGCGGCAGCGGCAGCGAGAGCAAGAGCACCGGCGGCAGCCAAGCCTATCTTCAACAATCTGGGGCTGAGCTTGTCCGGCCAGGAGCCTCCGTCAAAATGAGCTGCAAAACCTCAGGTTATACTTTTAGTAGCTATAACATGCATTGGGTAAAACAAACCCCCCGACAAGCATTGGAGTGGATAGGGGCCATATACCCCGGCAATGGAGACACAAGTTACAACCAGAAGTTTAAAGGCAAAGCTACACTCACAGTTGACAAATCCTCAAGTACTGCTTATATGCAACTCTCCTCTCTCACTTCCGAAGACAGTGCCGTATATTTTTGCACTCGGTCCAATTACTATGGATCTAGTGGCTGGTACTTTGACGTTTGGGGCACTGGGACAACTGTTACAGTG TCCAGCtrispecific Ab CD3-CD20 arm SEQ ID NO: 1483EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGKGGGGSGGGGSGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASLSVSSMHWYQQKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWIFNPPTFGGGTKLEIKGGSEGKSSGSGSESKSTGGSQAYLQQSGAELVRPGASVKMSCKTSGYTFSSYNMHWVKQTPRQALEWIGAIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCTRSNYYGSSGWYFDVWGTGTTVTVSS trispecific Ab CD3-CD20 arm SEQ ID NO: 1484GAGATCGTGCTGACCCAGTCTCCTGCCACACTGAGTGCTTCTCCAGGCGAGAGAGTGACCCTGTCCTGCTCCGCTTCCTCCTCCGTGTCCTACATGAACTGGTATCAGCAGAAGCCCGGCCAGGCTCCTCGGAGATGGATCTACGACTCTTCCAAGCTGGCCTCTGGTGTGCCAGCCAGATTTTCTGGCTCTGGCTCCGGCAGAGACTATACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTACTGCCAGCAGTGGTCTAGGAACCCTCCTACCTTTGGCGGAGGCACCAAGGTGGAAATCAAGGGCGGATCTGAGGGAAAGTCCAGCGGCTCCGGCAGCGAAAGCAAGTCCACCGGCGGAAGCCAGGTTCAACTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCTTCCGGCTACACCTTTACCAGATCCACCATGCACTGGGTCAAGCAGGCCCCTGGACAAGGCTTGGAGTGGATCGGCTACATCAACCCCAGCTCCGCCTACACCAACTACAACCAGAAATTCCAGGGCAGAGTGACCCTGACCGCCGACAAGTCTACCTCCACCGCCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTGCGCCTCTCCTCAGGTCCACTACGACTACGCCGGCTTTCCTTATTGGGGCCAGGGCACACTGGTCACCGTTTCTTCTGAGCCCAAATCTAGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTCCTGCGCCGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCGGTTCACGCAGAAGTCTCTCTCCCTGTCTCCGGGAAAAGGAGGCGGAGGATCTGGCGGAGGTGGAAGTGGCGGAGGCGGTTCTGGTGGTGGTGGATCTCAGATCGTGCTGTCTCAGTCTCCAGCTATCCTGTCTGCTAGCCCTGGCGAGAAAGTGACCATGACCTGTAGAGCCAGCCTGTCCGTGTCCTCCATGCACTGGTATCAGCAGAAGCCTGGCAGCTCCCCTAAGCCTTGGATCTACGCCACCTCCAATCTGGCCTCTGGCGTGCCAGCTAGATTCTCCGGATCTGGCTCCGGCACCTCCTACAGCCTGACAATCTCCAGAGTGGAAGCCGAGGATGCCGCCACCTACTACTGTCAGCAGTGGATCTTCAACCCTCCTACCTTCGGCGGAGGCACCAAGCTGGAAATCAAGGGAGGGAGCGAGGGAAAGTCCAGCGGAAGCGGCTCTGAGTCCAAATCCACCGGAGGGAGCCAGGCTTACTTGCAGCAGTCTGGTGCCGAACTCGTTAGACCTGGAGCCTCCGTGAAGATGTCCTGCAAGACCTCCGGCTACACCTTCTCCAGCTACAACATGCACTGGGTCAAGCAGACCCCTCGGCAGGCTCTGGAATGGATCGGCGCTATCTATCCTGGCAACGGCGACACCTCCTACAACCAGAAGTTCAAGGGCAAAGCTACCCTGACCGTGGACAAGTCCTCCTCCACCGCTTACATGCAGCTGTCCAGCCTGACCTCTGAGGACTCCGCCGTGTACTTCTGCACCCGGTCTAACTACTACGGCTCCTCCGGCTGGTACTTCGATGTGTGGGGAACCGGAACCACCGTGACAGTCTCTTCT trispecific Ab CD3-CD20 armSEQ ID NO: 1485 DIQMTQSPSSLSASVGDRVTITCRARQSIGTAIHWYQQKPGKAPKLLIKYASESISGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSGSWPYTFGQGTKLEIKGGSEGKSSGSGSESKSTGGSEVQLVESGGGLVKPGGSLRLSCAASGFTFSRYNMNWVRQAPGKGLEWVSSISTSSNYIYYADSVKGRFTFSRDNAKNSLDLQMSGLRAEDTAIYYCTRGWGPFDYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSGGGGSGGGGSEIVLTQSPATLSLSPGERATLSCRASLSVSSMHWYQQKPGQAPRLLIYATSNLASGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQWIFNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFSSYNMHWVRQAPGQGLEWMGAIYPGAGDTSYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARS NYYGSSGWYFDVWGKGTTVTVSStrispecific Ab CD3-CD20 arm SEQ ID NO: 1486GACATCCAGATGACCCAGTCTCCATCCTCTCTGTCCGCCTCTGTGGGCGACAGAGTGACCATTACCTGCCGGGCCAGACAGTCTATCGGCACCGCTATCCACTGGTATCAGCAGAAGCCTGGCAAGGCCCCTAAGCTGCTGATTAAGTACGCCTCCGAGTCCATCTCCGGCGTGCCCTCCAGATTTTCTGGCTCTGGATCTGGCACCGACTTTACCCTGACAATCTCCAGCCTGCAGCCTGAGGACTTCGCCACCTACTACTGTCAGCAGTCCGGCTCTTGGCCTTACACCTTTGGTCAGGGCACCAAGCTGGAAATCAAGGGCGGATCTGAGGGAAAGTCCAGCGGCTCCGGCAGCGAAAGCAAGTCCACCGGCGGAAGCGAGGTGCAGCTGGTTGAATCTGGCGGAGGACTGGTTAAGCCTGGCGGCTCTCTGAGACTGTCTTGTGCTGCTTCTGGCTTCACCTTCAGCCGGTACAACATGAACTGGGTCCGACAGGCTCCTGGCAAAGGCCTGGAATGGGTGTCCTCCATCTCCACCTCCAGCAACTACATCTACTACGCCGACTCCGTGAAGGGCAGATTCACCTTCTCCAGAGACAACGCCAAGAACTCCCTGGACCTGCAGATGTCTGGCCTGAGAGCTGAGGACACCGCTATCTACTACTGCACCAGAGGCTGGGGACCCTTCGATTATTGGGGCCAGGGAACCCTGGTCACCGTGTCATCTGAGCCCAAATCTAGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGTCTCTCTCCCTGTCTCCGGGAAAAGGAGGCGGAGGATCTGGCGGAGGTGGAAGTGGCGGAGGCGGTTCTGGTGGTGGTGGATCTGAGATCGTGCTGACCCAGTCTCCAGCCACACTGTCACTGTCTCCAGGCGAGAGAGCTACCCTGTCCTGTAGAGCCTCTCTGTCCGTGTCCTCCATGCACTGGTATCAGCAGAAGCCTGGACAGGCCCCTCGGCTGCTGATCTACGCTACCTCTAATCTGGCCAGCGGTATCCCCGCCAGATTTTCTGGTTCTGGCTCTGGCACCGACTTTACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTACTGCCAGCAGTGGATCTTCAACCCTCCTACCTTTGGCGGAGGCACCAAGGTGGAAATCAAGGGAGGGAGCGAGGGAAAGTCCAGCGGAAGCGGCTCTGAGTCCAAATCCACCGGAGGGAGCCAGGTTCAACTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCTCCTCCGTGAAGGTGTCCTGCAAGGCTTCCGGCTACACCTTCTCCAGCTACAACATGCACTGGGTCCGACAGGCCCCTGGACAAGGATTGGAATGGATGGGCGCTATCTACCCTGGCGCTGGCGATACCTCTTACGCCCAGAAATTCCAGGGCAGAGTGACCATCACCGCCGACGAGTCTACCTCCACCGCCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTGCGCCCGGTCTAATTACTACGGCTCCAGCGGCTGGTACTTCGACGTGTGGGGAAAG GGCACCACCGTGACAGTCTCTTCTtrispecific Ab CD3-CD20 arm SEQ ID NO: 1487EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGGGGSGGGGSGGGGSEIVLTQSPATLSLSPGERATLSCRASLSVSSMHWYQQKPGQAPRLLIYATSNLASGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQWIFNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFSSYNMHWVRQAPGQGLEWMGAIYPGAGDTSYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSNYYGSSGWYFDVWGKGTTVTVSS trispecific Ab CD3-CD20 arm SEQ ID NO: 1488GAGATCGTGCTGACCCAGTCTCCTGCCACACTGAGTGCTTCTCCAGGCGAGAGAGTGACCCTGTCCTGCTCCGCTTCCTCCTCCGTGTCCTACATGAACTGGTATCAGCAGAAGCCCGGCCAGGCTCCTCGGAGATGGATCTACGACTCTTCCAAGCTGGCCTCTGGTGTGCCAGCCAGATTTTCTGGCTCTGGCTCCGGCAGAGACTATACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTACTGCCAGCAGTGGTCTAGGAACCCTCCTACCTTTGGCGGAGGCACCAAGGTGGAAATCAAGGGCGGATCTGAGGGAAAGTCCAGCGGCTCCGGCAGCGAAAGCAAGTCCACCGGCGGAAGCCAGGTTCAACTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCTTCCGGCTACACCTTTACCAGATCCACCATGCACTGGGTCAAGCAGGCCCCTGGACAAGGCTTGGAGTGGATCGGCTACATCAACCCCAGCTCCGCCTACACCAACTACAACCAGAAATTCCAGGGCAGAGTGACCCTGACCGCCGACAAGTCTACCTCCACCGCCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTGCGCCTCTCCTCAGGTCCACTACGACTACGCCGGCTTTCCTTATTGGGGCCAGGGCACACTGGTCACCGTTTCTTCTGAGCCCAAATCTAGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGTCTCTCTCCCTGTCTCCGGGAAAAGGAGGCGGAGGATCTGGCGGAGGTGGAAGTGGCGGAGGCGGTTCTGGTGGTGGTGGATCTGAGATCGTGCTGACCCAGTCTCCAGCCACACTGTCACTGTCTCCAGGCGAGAGAGCTACCCTGTCCTGTAGAGCCTCTCTGTCCGTGTCCTCCATGCACTGGTATCAGCAGAAGCCTGGACAGGCCCCTCGGCTGCTGATCTACGCTACCTCTAATCTGGCCAGCGGTATCCCCGCCAGATTTTCTGGTTCTGGCTCTGGCACCGACTTTACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTACTGCCAGCAGTGGATCTTCAACCCTCCTACCTTTGGCGGAGGCACCAAGGTGGAAATCAAGGGAGGGAGCGAGGGAAAGTCCAGCGGAAGCGGCTCTGAGTCCAAATCCACCGGAGGGAGCCAGGTTCAACTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCTCCTCCGTGAAGGTGTCCTGCAAGGCTTCCGGCTACACCTTCTCCAGCTACAACATGCACTGGGTCCGACAGGCCCCTGGACAAGGATTGGAATGGATGGGCGCTATCTACCCTGGCGCTGGCGATACCTCTTACGCCCAGAAATTCCAGGGCAGAGTGACCATCACCGCCGACGAGTCTACCTCCACCGCCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTGCGCCCGGTCTAATTACTACGGCTCCAGCGGCTGGTACTTCGACGTGTGGGGAAAGGGCACCACCGTGACAGTCTCTTCTtrispecific/bispecific Ab CD79b arm HC SEQ ID NO: 1489QVQLQESGPGLVKPSETLSLTCSVSGASISSFYWSWIRQPADEGLEWIGRISPSGKTNYIPSLKSRIIMSLDASKNQFSLRLNSVTAADTAMYYCARGEYSGTYSYSFDVWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLS PGKtrispecific/bispecific Ab CD79b arm HC SEQ ID NO: 1490CAGGTTCAGCTGCAAGAGTCTGGTCCTGGCCTGGTCAAGCCTTCCGAGACACTGTCTCTGACCTGCTCTGTGTCCGGCGCCTCCATCTCTTCCTTCTACTGGTCCTGGATCCGGCAGCCTGCTGACGAAGGACTGGAATGGATCGGCCGGATCAGCCCTTCTGGCAAGACCAACTACATCCCCAGCCTGAAGTCCCGGATCATCATGTCCCTGGACGCCTCCAAGAACCAGTTCTCCCTGCGGCTGAACTCTGTGACCGCTGCCGATACCGCCATGTACTACTGTGCCAGAGGCGAGTACTCCGGCACCTACTCCTACAGCTTTGACGTGTGGGGACAAGGCACCATGGTCACAGTTTCTTCTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTCCTGCGCCGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCGGTTCACGCAGAAGTCTCTCTCCCTGTCTCCGG GAAAAtrispecific/bispecific Ab CD79b arm LC SEQ ID NO: 1491DIVMTQSPLSLSVTPGEPASISCRSSESLLDSEDGNTYLDWFLQKPGQSPQLLIYTLSYRASGVPDRFSGSGSDTDFTLHISSLEAEDVGLYYCMQRMEFPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECtrispecific/bispecific Ab CD79b arm LC SEQ ID NO: 1492GACATCGTGATGACCCAGTCTCCACTGAGCCTGTCTGTGACACCTGGCGAGCCTGCCTCCATCTCCTGTAGATCTTCTGAGTCCCTGCTGGACAGCGAGGACGGCAATACCTACCTGGACTGGTTCCTGCAGAAGCCCGGACAGTCTCCTCAGCTGCTGATCTACACCCTGTCCTACAGAGCCTCTGGCGTGCCCGATAGATTCTCCGGCTCTGGCTCTGACACCGACTTTACCCTGCACATCTCCAGCCTGGAAGCCGAGGATGTGGGCCTGTACTACTGTATGCAGCGGATGGAATTTCCCCTGACCTTCGGCCAGGGCACCAAGGTGGAAATCAAGCGCACCGTGGCCGCCCCTAGCGTGTTTATCTTCCCTCCCTCGGATGAGCAGCTTAAGTCAGGCACCGCATCCGTGGTCTGCCTGCTCAACAACTTCTACCCGAGGGAAGCCAAAGTGCAGTGGAAAGTGGACAACGCGCTCCAGTCGGGAAACTCCCAGGAGTCCGTGACCGAACAGGACTCCAAGGACAGCACTTATTCCCTGTCCTCCACTCTGACGCTGTCAAAGGCCGACTACGAGAAGCACAAGGTCTACGCCTGCGAAGTGACCCATCAGGGGCTTTCCTCGCCCGTGACTAAGAGCTTCAATCGGGGCGAATGC trispecific/bispecific Ab CD79b arm HCSEQ ID NO: 1493 QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSATWNWIRQSPSRGLEWLGRTYYRSKWYNDYTVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCTRVDIAFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPG Ktrispecific/bispecific Ab CD79b arm HC SEQ ID NO: 1494CAGGTTCAGCTGCAGCAGTCTGGCCCTGGACTGGTCAAGCCCTCTCAGACCCTGTCTCTGACCTGTGCCATCTCCGGCGACTCCGTGTCCTCTAATTCTGCCACCTGGAACTGGATCCGGCAGTCCCCTAGTAGAGGCCTGGAATGGCTGGGCAGAACCTACTACCGGTCCAAGTGGTACAACGACTACACCGTGTCCGTGAAGTCCCGGATCACCATCAATCCCGACACCTCCAAGAACCAGTTCTCCCTGCAGCTCAACAGCGTGACCCCTGAGGATACCGCCGTGTACTACTGCACCAGAGTGGATATCGCCTTCGACTACTGGGGCCAGGGCACACTGGTTACCGTTTCTTCTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTCCTGCGCCGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCGGTTCACGCAGAAGTCTCTCTCCCTGTCTCCGGGAAAAtrispecific/bispecific Ab CD79b arm LC SEQ ID NO: 1495QTVVTQPPSVSEAPRQRVTISCSGSSSNIGNHGVNWYQQLPGKAPKLLIYNDDLLPSGVSDRFSGSTSGTSGSLAISGLQSEDEADYYCAAWDDSLNGVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECStrispecific/bispecific Ab CD79b arm LC SEQ ID NO: 1496CAGACAGTGGTCACCCAGCCTCCATCTGTGTCTGAGGCCCCTAGACAGAGAGTGACCATCTCCTGCTCCGGCTCCTCCTCCAACATCGGCAATCATGGCGTGAACTGGTATCAGCAGCTGCCCGGCAAGGCTCCCAAACTGCTGATCTACAACGACGACCTGCTGCCTTCTGGCGTGTCCGACAGATTCTCCGGCTCTACCTCTGGCACCTCTGGATCCCTGGCTATCTCTGGCCTGCAGTCTGAGGACGAGGCCGACTACTATTGTGCCGCCTGGGACGATTCTCTGAACGGCGTTGTGTTTGGCGGAGGCACCAAGCTGACAGTGTTGGGACAGCCTAAGGCAGCCCCCTCCGTGACCCTGTTCCCGCCATCATCCGAAGAACTGCAGGCCAACAAGGCCACGCTCGTGTGCCTGATTTCCGACTTCTACCCGGGGGCCGTGACTGTGGCCTGGAAGGCAGACTCAAGCCCTGTGAAGGCTGGCGTCGAGACTACCACCCCGTCGAAGCAATCCAACAACAAATACGCGGCGTCCAGCTACCTGAGCCTGACCCCTGAGCAGTGGAAATCCCACCGGTCCTACTCGTGCCAAGTCACCCACGAGGGATCCACTGTGGAAAAGACCG TGGCGCCGACTGAGTGTTCCtrispecific/bispecific Ab CD79b arm HC SEQ ID NO: 1497QVQLQESGPGLVKPSQTLSLTCTVSGVSISNYYWSWIRQPPGKGLEWIGRISPSGRTNYNPSLKSRVTMSLDASKNQFSLKLSSVTAADTAVYYCARGEYSGTYSYSFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLS PGKtrispecific/bispecific Ab CD79b arm HC SEQ ID NO: 1498CAGGTTCAGCTGCAAGAGTCTGGCCCTGGCCTGGTCAAGCCCTCTCAGACCCTGTCTCTGACCTGTACCGTGTCCGGCGTGTCCATCTCCAACTACTACTGGTCCTGGATCCGGCAGCCTCCTGGCAAAGGACTGGAATGGATCGGCCGCATCTCTCCTTCTGGTCGCACCAACTACAACCCCAGCCTGAAAAGCAGAGTGACCATGTCTCTGGACGCCTCCAAGAACCAGTTCTCCCTGAAGCTGTCCTCCGTGACCGCTGCTGATACCGCCGTGTACTACTGTGCCAGAGGCGAGTACTCCGGCACCTACTCCTACAGCTTCGACATCTGGGGCCAGGGCACCATGGTCACAGTCTCTTCTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTCCTGCGCCGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCGGTTCACGCAGAAGTCTCTCTCCCTGTCTCCGG GAAAAtrispecific/bispecific Ab CD79b arm LC SEQ ID NO: 1499DIQMTQSPSSLSASVGDRVTITCRSSQSLFDSDDGNTYLDWFQQKPGQSPKLLIQTLSYRASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCMQRMEFPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECtrispecific/bispecific Ab CD79b arm LC SEQ ID NO: 1500GACATCCAGATGACCCAGTCTCCATCCTCTCTGTCCGCCTCTGTGGGCGACAGAGTGACCATCACCTGTCGGTCCTCTCAGTCCCTGTTCGACTCTGACGACGGCAACACCTACCTGGACTGGTTCCAGCAGAAGCCCGGCCAGTCTCCTAAGCTGCTGATCCAGACACTGTCCTACAGAGCCTCTGGCGTGCCCTCCAGATTTTCCGGCTCTGGCTCTGGCACCGACTTTACCCTGACAATCTCCAGCCTGCAGCCTGAGGACTTCGCCACCTACTACTGTATGCAGCGGATGGAATTTCCCCTGACCTTCGGCGGAGGCACCAAGGTGGAAATCAAGCGCACCGTGGCCGCCCCTAGCGTGTTTATCTTCCCTCCCTCGGATGAGCAGCTTAAGTCAGGCACCGCATCCGTGGTCTGCCTGCTCAACAACTTCTACCCGAGGGAAGCCAAAGTGCAGTGGAAAGTGGACAACGCGCTCCAGTCGGGAAACTCCCAGGAGTCCGTGACCGAACAGGACTCCAAGGACAGCACTTATTCCCTGTCCTCCACTCTGACGCTGTCAAAGGCCGACTACGAGAAGCACAAGGTCTACGCCTGCGAAGTGACCCATCAGGGGCTTTCCTCGCCCGTGACTAAGAGCTTCAATCGGGGCGAATGC trispecific/bispecific Ab CD79b arm LCSEQ ID NO: 1501 GACATCCAGATGACCCAGAGCCCTAGCAGCCTGAGCGCCAGCGTGGGCGACAGAGTGACCATTACCTGCAGAAGCAGCCAGAGCCTGTTCGACAGCGACGACGGCAATACCTACCTGGACTGGTTCCAGCAGAAGCCTGGCCAGAGCCCTAAGCTGCTGATCCAGACCCTGAGCTACAGAGCCAGCGGCGTGCCTAGCAGATTCTCCGGCAGCGGCTCCGGCACCGACTTCACCCTGACCATCAGCAGCCTGCAGCCTGAGGACTTCGCCACCTACTACTGCATGCAGAGAATGGAGTTCCCTCTGACCTTCGGCGGCGGCACCAAGGTGGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT trispecific/bispecific Ab CD79b arm HCSEQ ID NO: 1502 QVQLQESGPGLVKPSQTLSLTCTVSGVSISNYYWSWIRQPPGKGLEWIGRISPSGRTNYNPSLKSRVTMSLDASKNQFSLKLSSVTAADTAVYYCARGEYSGTYSYSFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGKtrispecific/bispecific Ab CD79b arm HC SEQ ID NO: 1503CAGGTTCAGCTGCAAGAGTCTGGCCCTGGCCTGGTCAAGCCCTCTCAGACCCTGTCTCTGACCTGTACCGTGTCCGGCGTGTCCATCTCCAACTACTACTGGTCCTGGATCCGGCAGCCTCCTGGCAAAGGACTGGAATGGATCGGCCGCATCTCTCCTTCTGGTCGCACCAACTACAACCCCAGCCTGAAAAGCAGAGTGACCATGTCTCTGGACGCCTCCAAGAACCAGTTCTCCCTGAAGCTGTCCTCCGTGACCGCTGCTGATACCGCCGTGTACTACTGTGCCAGAGGCGAGTACTCCGGCACCTACTCCTACAGCTTCGACATCTGGGGCCAGGGCACCATGGTCACAGTCTCTTCTGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGTCTCTCTCCCTGTCTCCGG GAAAAtrispecific/bispecific Ab CD79b arm LC SEQ ID NO: 1499DIQMTQSPSSLSASVGDRVTITCRSSQSLFDSDDGNTYLDWFQQKPGQSPKLLIQTLSYRASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCMQRMEFPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

Table 32 provides a summary of examples of some CD79b×CD3 bispecificantibodies described herein.

TABLE 32 Exemplary CD79b × CD3 bispecific antibodies HC1 LC HC2 aminoHC1 amino amino HC2 acid DNA acid LC DNA acid DNA HC1/LC sequencesequence sequence sequence sequence sequence (CD79b SEQ ID SEQ ID SEQ IDSEQ ID HC2 SEQ ID SEQ ID ID arm) NO NO NO NO (CD3-arm) NO NO 79C3B601CD9B374 1489 1490 1491 1492 CD3B2030- 1504 1505 N106A 79C3B646 CD9B330-1493 1494 1495 1496 CD3B2030- 1504 1505 N31S N106A 79C3B651 CD9B643 14971498 1499 1500 CD3B2030- 1504 1505 N106A 79C3B605 CD9B374 1489 1490 14911492 CD3B2089- 1506 1507 N106G 79C3B645 CD9B330- 1493 1494 1495 1496CD3B2089- 1506 1507 N31S N106G 79C3B650 CD9B643 1497 1498 1499 1500CD3B2089- 1506 1507 N106G

bispecific Ab CD3-arm SEQ ID NO: 1504EIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK bispecific Ab CD3-armSEQ ID NO: 1505GAGATCGTGCTGACCCAGTCTCCTGCCACACTGAGTGCTTCTCCAGGCGAGAGAGTGACCCTGTCCTGCTCCGCTTCCTCCTCCGTGTCCTACATGAACTGGTATCAGCAGAAGCCCGGCCAGGCTCCTCGGAGATGGATCTACGACTCTTCCAAGCTGGCCTCTGGTGTGCCAGCCAGATTTTCTGGCTCTGGCTCCGGCAGAGACTATACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTACTGCCAGCAGTGGTCTAGGAACCCTCCTACCTTTGGCGGAGGCACCAAGGTGGAAATCAAGGGCGGATCTGAGGGAAAGTCCAGCGGCTCCGGCAGCGAAAGCAAGTCCACCGGCGGAAGCCAGGTTCAACTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCTTCCGGCTACACCTTTACCAGATCCACCATGCACTGGGTCAAGCAGGCCCCTGGACAAGGCTTGGAGTGGATCGGCTACATCAACCCCAGCTCCGCCTACACCAACTACAACCAGAAATTCCAGGGCAGAGTGACCCTGACCGCCGACAAGTCTACCTCCACCGCCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTGCGCCTCTCCTCAGGTCCACTACGACTACGCCGGCTTTCCTTATTGGGGCCAGGGCACACTGGTCACCGTTTCTTCTGAGCCCAAATCTAGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGTCTCTCTCCCTGTCTCCGGGAAAASEQ ID NO: 1506 bispecific Ab CD3 armEIVLTQSPATLSASPGERVTLSCSASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGSGRDYTLTISSLEPEDFAVYYCQQWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQSGAEVKKPGSSVKVSCKASGYTFTRSTMHWVRQAPGQGLEWMGYINPSSAYTNYAQKFQGRVTLTADKSTSTAYMELSSLRSEDTAVYYCASPQVHYDYGGFPYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK bispecific Ab CD3 armSEQ ID NO: 1507GAGATCGTGCTGACCCAGTCTCCTGCCACACTGAGTGCTTCTCCAGGCGAGAGAGTGACCCTGTCCTGCTCCGCTTCCTCCTCCGTGTCCTACATGAACTGGTATCAGCAGAAGCCCGGCCAGGCTCCTCGGAGATGGATCTACGACTCTTCCAAGCTGGCCTCTGGTGTGCCAGCCAGATTTTCTGGCTCTGGCTCCGGCAGAGACTATACCCTGACCATCTCCAGCCTGGAACCTGAGGACTTCGCCGTGTACTACTGCCAGCAGTGGTCTAGGAACCCTCCTACCTTTGGCGGAGGCACCAAGGTGGAAATCAAGGGCGGATCTGAGGGAAAGTCCAGCGGCTCCGGCAGCGAAAGCAAGTCCACCGGCGGAAGCCAGGTTCAACTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCTCCTCCGTGAAAGTGTCCTGCAAGGCTTCCGGCTACACTTTTACCAGATCCACCATGCACTGGGTCCGACAGGCTCCAGGACAAGGCTTGGAGTGGATGGGCTACATCAACCCCAGCTCCGCCTACACCAACTACGCCCAGAAATTCCAGGGCAGAGTGACCCTGACCGCCGACAAGTCTACCTCCACCGCCTACATGGAACTGTCCAGCCTGAGATCTGAGGACACCGCCGTGTACTACTGCGCTTCTCCTCAGGTGCACTACGACTACGGCGGCTTTCCTTATTGGGGCCAGGGCACACTGGTCACCGTTTCTTCTGAGCCCAAATCTAGCGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCCGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGAGCGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTGTCGAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGATGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGTCTCTCTCCCTGTCTCCGGGAAAA

The antibodies were expressed in ExpiCHO-S™ cells (ThermoFisherScientific; Waltham, Mass., Cat #A29127) by transient transfection withpurified plasmid DNA following the manufacturer's recommendations.Briefly, ExpiCHO-S™ cells were maintained in suspension in ExpiCHO™expression medium (ThermoFisher Scientific, Cat #A29100) in an orbitalshaking incubator set at 37° C., 8% CO₂ and 125 RPM. The cells werepassaged and diluted prior to transfection to 6.0×10⁶ cells per ml,maintaining cell viability at 99.0% or better. Transient transfectionswere done using the ExpiFectamine™ CHO transfection kit (ThermoFisherScientific, Cat #A29131). For each ml of diluted cells to betransfected, 0.5 microgram of bispecific encoding DNA (HC1:HC2:LC=1:2:2)and 0.5 microgram of pAdVAntage DNA (Promega, Cat #E1711) was used anddiluted into OptiPRO™ SFM complexation medium. ExpiFectamine™ CHOreagent was used at a 1:4 ratio (v/v, DNA:reagent) and diluted intoOptiPRO™. The diluted DNA and transfection reagent were combined for oneminute, allowing DNA/lipid complex formation, and then added to thecells. After overnight incubation, ExpiCHO™ feed and ExpiFectamine™ CHOenhancers were added to the cells as per the manufacturer's Standardprotocol. Cells were incubated with orbital shaking (125 rpm) at 37° C.for seven days prior to harvesting the culture broth. The culturesupernatant from the transiently transfected ExpiCHO-S™ cells wasclarified by centrifugation (30 min, 3000 rcf) followed by filtration(0.2 μm PES membrane, Corning; Corning, N.Y.).

The filtered cell culture supernatant was loaded onto a pre-equilibrated(1×DPBS, pH 7.2) Mab Select Sure Protein A column (GE Healthcare) usingan AKTAXpress chromatography system. After loading, the column waswashed with 10 column volumes of 1×DPBS, pH7.2. The protein was elutedwith 10 column volumes of 0.1 M sodium (Na)-Acetate, pH 3.5. Proteinfractions were neutralized immediately by the addition of 2.5 M TrisHCl, pH 7.2 to 20% (v/v) of the elution fraction volume. Peak fractionswere pooled and loaded onto a CH1 column (Thermofisher). After loading,the column was washed with 10 column volumes of 1×DPBS, pH7.2. Theprotein was eluted with 10 column volumes of 0.1 M sodium (Na)-Acetate,pH 3.5. Protein fractions were partially neutralized by the addition of2.5 M Tris HCl, pH 7.2 to 15% (v/v) of the final volume. The highmolecular weight species were removed by preparative size exclusionchromatography (SEC) using Superdex 200 (GE Healthcare). Post sampleinjection, the column was developed with 1×DPBS and the major peakfractions were pooled, dialyzed into 10 mM Histidine, pH6.5 and filtered(0.2 μm).

The concentration of purified protein was determined by absorbance at280 nm on a Dropsense spectrophotometer. The quality of the purifiedprotein was assessed by cSDS and analytical size exclusion HPLC (AgilentHPLC system). The endotoxin level was measured using a turbidometric LALassay (Pyrotell®-T, Associates of Cape Cod; Falmouth, Mass.).

Example 4: Bispecific and Trispecific Antibodies BindingCharacterization

Binding of Bispecific CD79×CD3 Antibodies on CD79⁺ Target Cells

The binding affinity of the CD79b binding arm of the CD79×CD3 bispecificmolecules were assessed using cell lines that were validated by flowcytometry to have different endogenous expression levels of CD79b on thecell surface, shown in Table 33.

TABLE 33 CD79b Antigen Density of B Lymphoma Cell Lines CD79b AntigenDensity (Antigen Cell Line Cell Type Number/cell) HBL-1 Diffuse largeB-cell lymphoma line 429,649 OCI-LY-10 Diffuse large B-cell lymphomaline 38,885 CARNAVAL Diffuse large B-cell lymphoma line 98,176 WILL2Diffuse large B-cell lymphoma line 3,824

Diffuse large B-cell lymphoma cell lines were incubated for 1 hour withCD79b×CD3 test molecules 79C3B646, 79C3B651, and 79C3B601 (1 uM startingconcentration at 1:3 serial dilutions) at 37° C. All cells were washedwith BD stain buffer (BD Biosciences; Cat #554657), centrifuged at 1200RPM for 3 mins, with supernatant discarded. Cells were then stained for20 minutes at 4° C. with BD stain buffer containing AlexaFluor 647labeled anti-human IgG secondary antibody (Jackson Immuno; Cat#109-606-098) at a 1:200 dilution along with Aqua Fixable Live/Deadstain (Invitrogen; Cat #L34957) at a 1:400 dilution. All cells werewashed with BD stain buffer (BD Biosciences; Cat #554657), centrifugedat 1200 RPM for 3 minutes, with supernatant discarded. Cells wereanalyzed using Intellicyt (Sartorius) flow cytometer and meanfluorescent intensity (MFI) was generated using Forcyt software(Sartorius). MFI was graphed and EC50 values generated using GraphPadPRISM v.8. Dose response curves were generated by transforming the xaxis values using the formula x=lox. Data were then graphed usingnon-linear regression curve fit analysis “log(agonist) vs.response-variable slope (four parameter)”.

All CD79b×CD3 molecules showed good binding on cell lines expressingendogenous CD79b on the cell surface, with the CD79b binding arm ofconstruct 79C3B651 showing the highest binding affinity across alltested cell lines, shown in FIGS. 7A-7D and Table 34.

TABLE 34 CD79b × CD3 Bispecifics Cell Binding EC50 Values HBL-1 OCI-LY10Carnaval WILL-2 EC50 EC50 EC50 EC50 (nM) (nM) (nM) (nM) 79C3B646 97undetermined 44 undetermined 79C3B651 15 undetermined 12 undetermined79C3B601 48 undetermined 89 undetermined

Binding of Trispecific CD79×CD20×CD3 Antibodies on CD79b⁺ and CD20⁺Target Cells

The binding affinity of the CD79b binding arm of the CD79×CD20×CD3trispecific molecules as well as control CD79b×CD3 and Null×CD20×CD3were assessed using cell lines that were validated by flow cytometry tohave different endogenous expression levels of CD79b and CD20 on thecell surface, shown in Table 35.

TABLE 35 CD79b and CD20 Antigen Density of B Lymphoma Cell Lines CD20Antigen CD79b Antigen Density Density (Antigen (Antigen Cell Line CellType Number/cell) Number/cell) HBL-1 Diffuse large B-cell 429,649 73,467lymphoma line OCI-LY-10 Diffuse large B-cell 38,885 67,352 lymphoma lineCARNAVAL Diffuse large B-cell 98,176 118,789 lymphoma line WILL2 Diffuselarge B-cell 3,824 314 lymphoma line

Diffuse large B-cell lymphoma cell lines were incubated for 1 hour withCD79b×CD20×CD3 test molecules C923B74, C923B99, and C923B38; CD79×CD3test molecules 79C3B646, 79C3B651, and 79C3B601 and Null×CD20×CD3control molecule C923B98 (1 μM starting concentration at 1:3 serialdilutions) at 37° C. All cells were washed with BD stain buffer (BDBiosciences; Cat #554657), centrifuged at 1200 RPM for 3 minutes, withsupernatant discarded. Cells were then stained for 20 minutes at 4° C.with BD stain buffer containing AlexaFluor 647 labeled anti-human IgGsecondary antibody (Jackson Immuno; Cat #109-606-098) at a 1:200dilution along with Aqua Fixable Live/Dead stain (Invitrogen; Cat#L34957) at a 1:400 dilution. All cells were washed with BD stain buffer(BD Biosciences; Cat #554657), centrifuged at 1200 RPM for 3 mins, withsupernatant discarded. Cells were analyzed using Intellicyt (Sartorius)flow cytometer and mean fluorescent intensity (MFI) was generated usingForcyt software (Sartorius). MFI was graphed and EC50 values generatedusing GraphPad PRISM v.8. Dose response curves were generated bytransforming the x axis values using the formula x=lox. Data was thengraphed using non-linear regression curve fit analysis “log(agonist) vs.response-variable slope (four parameter)”.

All CD79b×CD20×CD3 molecules showed good binding on cell linesexpressing endogenous CD79b and CD20 on the cell surface, with sometrispecific constructs showing better binding affinity across cell lineswhen compared to binding of CD79b×CD3 and CD20×CD3 control molecules,shown in FIGS. 8A-8D and Table A-10. The CD79b binding arm oftrispecific construct C923B99 showed the highest binding affinity acrossall tested cell lines, shown in FIGS. 8A-8D and Table 36.

TABLE 36 CD79b × CD20 × CD3 Trispecific Cell Binding EC50 Values HBL-1OCI-LY10 Carnaval EC50 EC50 EC50 WILL-2 EC50 (nM) (nM) (nM) (nM) C923B3843 12 16 undetermined C923B74 52 66 23 undetermined C923B99 8 2 6undetermined 79C3B646 97 undetermined 44 undetermined 79C3B651 15undetermined 12 undetermined 79C3B601 48 undetermined 89 undeterminedC923B98 undetermined undetermined Undetermined undetermined

Kinetic Cell Binding of Bispecific CD79×CD3 Antibodies on CD79⁺ TargetCells

The binding kinetics of the CD79b binding arm of the CD79×CD3 bispecificmolecules were assessed over a time course using cell lines that werevalidated by flow cytometry to have different endogenous expressionlevels of CD79b on the cell surface, shown in Table 37.

TABLE 37 CD79b Antigen Density of B Lymphoma Cell Lines CD79b AntigenDensity (Antigen Cell Line Cell Type Number/cell) HBL-1 Diffuse largeB-cell 429,649 lymphoma line OCI-LY10 Diffuse large B-cell 38,885lymphoma line CARNAVAL Diffuse large B-cell 98,176 lymphoma line

Diffuse large B-cell lymphoma cell lines were incubated for 1, 3, 24,and 48 hours with CD79b×CD3 test molecules 79C3B646, 79C3B651, and79C3B601 (300 nM, 60 nM, 12 nM) at 37° C. At each time point, cells werewashed with BD stain buffer (BD Biosciences; Cat #554657), centrifugedat 1200 RPM for 3 mins, with supernatant discarded. Cells were thenstained for 30 minutes at 4° C. with BD stain buffer containingAlexaFluor 647 labeled anti-human IgG secondary antibody (JacksonImmuno; Cat #109-606-098) at a 1:200 dilution. All cells were washedwith BD stain buffer (BD Biosciences; Cat #554657), centrifuged at 1200RPM for 3 mins, with supernatant discarded. Cells were resuspended in50ul of FACS buffer containing a 1:1000 dilution of Cytox Greenviability dye (Invitrogen, Cat #S34860). Cells were analyzed usingIntellicyt (Sartorius) flow cytometer and mean fluorescent intensity(MFI) was generated using Forcyt software (Sartorius). MFI was graphedand EC50 values generated using GraphPad PRISM v.8.

All CD79b×CD3 bispecific constructs showed steady CD79b binding kineticswith minimal loss of signal over time, as shown in FIGS. 9A-9I. 79C3B651showed superior binding kinetics and the least amount of signal lossover time, shown in FIGS. 9A-9I.

Kinetic Cell Binding of Trispecific CD79×CD20×CD3 Antibodies on CD79b⁺and CD20⁺ Target Cells

The binding kinetics of the CD79b and CD20 binding arms of theCD79×CD20×CD3 trispecific molecules were assessed over a time courseusing cell lines that were validated by flow cytometry to have differentendogenous expression levels of CD79b and CD20 on the cell surface,shown in Table 38.

TABLE 38 CD79b and CD20 Antigen Density of B Lymphoma Cell Lines CD79bAntigen CD20 Antigen Density (Antigen Density (Antigen Cell Line CellType Number/cell) Number/cell) HBL-1 Diffuse large B-cell 429,649 73,467lymphoma line OCI-LY-10 Diffuse large B-cell 38,885 67,352 lymphoma lineCARNAVAL Diffuse large B-cell 98,176 118,789 lymphoma line

Diffuse large B-cell lymphoma cell lines were incubated for 1, 3, 24,and 48 hours with CD79b×CD20×CD3 test molecules C923B74, C923B99, andC923B38; CD79×CD3 test molecules 79C3B646, 79C3B651, and 79C3B601 andNull×CD20×CD3 control molecule C923B98 (300 nM, 60 nM, 12 nM) at 37° C.At each time point, cells were washed with BD stain buffer (BDBiosciences; Cat #554657), centrifuged at 1200 RPM for 3 mins, withsupernatant discarded. Cells were then stained for 30 minutes at 4° C.with BD stain buffer containing AlexaFluor 647 labeled anti-human IgGsecondary antibody (Jackson Immuno; Cat #109-606-098) at a 1:200dilution. All cells were washed with BD stain buffer (BD Biosciences;Cat #554657), centrifuged at 1200 RPM for 3 mins, with supernatantdiscarded. Cells were resuspended in 50ul of FACS buffer containing a1:1000 dilution of Cytox Green viability dye (Invitrogen, Cat #S34860).Cells were analyzed using Intellicyt (Sartorius) flow cytometer and meanfluorescent intensity (MFI) was generated using Forcyt software(Sartorius). MFI was graphed and EC50 values generated using GraphPadPRISM v.8.

All CD79b×CD20×CD3 bispecific constructs showed steady CD79b bindingkinetics with minimal loss of signal over time, shown in FIGS. 10A-10I.Trispecific construct C923B99 and bispecific construct 79C3B651, whichboth have the same CD79b and CD20 binding arms, showed superior bindingkinetics and the least amount of signal loss over time, shown in FIGS.10A-10I.

Binding of Bispecific CD79×CD3 Antibodies and Trispecific CD79×CD20×CD3Antibodies on Pan T-Cells

Binding of the CD3 arm of CD79×CD3 bispecific and CD79b×CD20×CD3trispecific constructs was assessed using cryo-preserved, negativelyselected, primary human CD3⁺ pan T cells. Primary human CD3⁺ pan T cellsfrom four different donors were incubated for 1 hour with CD79b×CD20×CD3test molecules C923B74, C923B99, and C923B38 or CD79×CD3 test molecules79C3B646, 79C3B651 (1 uM starting concentration at 1:3 serial dilutions)at 37° C. All cells were washed with BD stain buffer (BD Biosciences;Cat #554657), centrifuged at 1200 RPM for 3 mins, with supernatantdiscarded. Cells were then stained for 20 minutes at 4° C. with BD stainbuffer containing AlexaFluor 647 labeled anti-human IgG secondaryantibody (Jackson Immuno; Cat #109-606-098) at a 1:300 dilution. Allcells were washed with BD stain buffer (BD Biosciences; Cat #554657),centrifuged at 1200 RPM for 3 mins, with supernatant discarded. Cellswere resuspended in 50ul of FACS buffer containing a 1:1000 dilution ofCytox Green viability dye (Invitrogen, Cat #534860). Cells were analyzedusing Intellicyt (Sartorius) flow cytometer and mean fluorescentintensity (MFI) was generated using Forcyt software (Sartorius). MFI wasgraphed using GraphPad PRISM v.8. Dose response curves were generated bytransforming the x axis values using the formula x=lox. Data was thengraphed using non-linear regression curve fit analysis “log(agonist) vs.response-variable slope (four parameter)”.

All CD79b×CD20×CD3 and CD79b×CD3 molecules showed moderate binding onall donor Pan T cells expressing endogenous CD3 on the cell surface,shown in FIGS. 11A-11D.

Example 5: Functional Characterization: Antagonistic Activity ofCD79×CD3 Bispecific and CD79×CD20×CD3 Trispecific Antibodies

Bispecific CD79×CD3 and Trispecific CD79×CD20×CD3 Mediated CytotoxicityAgainst CD79b⁺ and CD79B⁻ Target Cells

mKATE2 DLBCL target cells were maintained in complete RPMI(ThermoFisher, catalog #11875093)1640 media containing 10% heatinactivated fetal bovine serum. Prior to the assay, antibodies were madeat 3-fold serial dilutions in the at RPMI 1640 media containing 10% heatinactivated fetal bovine serum, at 4-fold expected final concentration.A volume of 50 μL of medium-diluted bsAb or trispecific Ab in each wellof a 96-well plate were further diluted into 200 μL by adding a mix oftarget and effector cell suspension. The target cell lines wereharvested by centrifuge at 400×g for 5 min, washed one time with phenolred-free RPMI 1640 media, counted and suspended in fresh complete phenolred-free RPMI 1640 media at 1×10⁶ cells/mL. Healthy donor T cells(isolated by CD3—negative selection provided by Discovery Life Sciences)were thawed in complete phenol red-free media (RPMI 1640 mediacontaining 10% heat inactivated fetal bovine serum), counted andsuspended in fresh complete phenol red-free RPMI 1640 media at 1×10⁶cells/mL. Target cells and T cells were mixed to obtain 5:1 effector totarget cell ratio. Cell suspension was added to antibody dilution wellsaccording to plate layout (150 μL/well).

After mixing target and T cells with corresponding bsAb dilution, 80 μLfrom each well, containing 200 μl with 10000 target and 50000 T cells,were dispensed in a 384 well plate, in duplicate. Plates were sealedusing a Breathe-Easy membrane seal. Next, co-cultures were placed in anIncuCyte ZOOM live-content imaging system, and images were automaticallyacquired in both phase and fluorescence channels every 6 hours for 3 to6 days with a 4× objective lens (single whole well image). IncuCyte Zoomsoftware was used to detect target cells based on mKATE2 expressionusing optimized process definition parameters. To measure the amount oftarget cells/well, the total red area was quantified, and raw valueswere exported in Excel (Microsoft Office). To quantify cancer cellkilling over time, the average values for each replicate were pasted inPrism (GraphPad; version 7 for PC). Expansion indexes (EI) per timepointwere calculated by dividing value at Tx by TO. Growth inhibition (GI)was calculated by normalizing each timepoint to the value of theuntreated well average at that timepoint. From the GI values, area underthe curve (AUC) values were derived for each condition. Afternormalizing the AUC to the untreated control (target with effector),antibody concentrations were plotted against the AUC values as a doseresponse. EC50 values were generated using GraphPad PRISM v.8. Doseresponse curves were generated by transforming the x axis values usingthe formula x=lox. Data was then graphed using non-linear regressioncurve fit analysis “log(agonist) vs. response-variable slope (fourparameter)”. Lead CD79b×CD3 bispecific antibodies and CD79b×CD20×CD3trispecific antibodies (79C3B646, C923B74, 79C3B601, C923B38, 79C3B651,C923B99, 79C3B613, C923B98) were evaluated for cytotoxicity on HBL1 andOCI-Ly10 cells. IC50 (pM) values are listed in Table 39, Table 40, Table41, and Table 42.

TABLE 39 HBL-1 killing Incucyte (Average of 2 independent experiments)Protein ID CD79b CD20 CD3 IC50 (pM) 79C3B645 CD9B330 NA CD3B2089 7189.079C3B646 CD9B330 NA CD3B2030 257.4 C923B73 CD9B330 C20B22 CD3B20896805.0 C923B74 CD9B330 C20B22 CD3B2030 346.3 79C3B605 CD9B374 NACD3B2089 29549.0 79C3B601 CD9B374 NA CD3B2030 203.9 C923B36 CD9B374C20B22 CD3B2089 31040.0 C923B38 CD9B374 C20B22 CD3B2030 301.2 79C3B650CD9B643 NA CD3B2089 43314.0 79C3B651 CD9B643 NA CD3B2030 32.5 C923B95CD9B643 C20B22 CD3B2089 4891.0 C923B99 CD9B643 C20B22 CD3B2030 69.2

TABLE 40 OCI-Ly10 killing Incucyte (Average of 2 independentexperiments) Protein ID CD79b CD20 CD3 IC50 (nM) 79C3B645 CD9B330 NACD3B2089 18.0 79C3B646 CD9B330 NA CD3B2030 18.3 C923B73 CD9B330 C20B22CD3B2089 132.4 C923B74 CD9B330 C20B22 CD3B2030 25.6 79C3B605 CD9B374 NACD3B2089 54.3 79C3B601 CD9B374 NA CD3B2030 11.7 C923B36 CD9B374 C20B22CD3B2089 42.0 C923B38 CD9B374 C20B22 CD3B2030 8.0 79C3B650 CD9B643 NACD3B2089 7.0 79C3B651 CD9B643 NA CD3B2030 4.7 C923B95 CD9B643 C20B22CD3B2089 14.8 C923B99 CD9B643 C20B22 CD3B2030 5.6

TABLE 41 CARNAVAL killing (Incucyte) Protein ID CD79b CD20 CD3 IC50 (nM)79C3B646 CD9B330 NA CD3B2030 1.393 C923B74 CD9B330 C20B22 CD3B2030 0.74179C3B601 CD9B374 NA CD3B2030 1.645 C923B38 CD9B374 C20B22 CD3B2030 0.465C923B99 CD9B643 C20B22 CD3B2030 0.285

TABLE 42 Daudi killing (Incucyte) Protein ID CD79b CD20 CD3 IC50 (nM)79C3B646 CD9B330 NA CD3B2030 0.597 C923B74 CD9B330 C20B22 CD3B2030 0.10079C3B601 CD9B374 NA CD3B2030 0.406 C923B38 CD9B374 C20B22 CD3B2030 0.071C923B99 CD9B643 C20B22 CD3B2030 < Cone tested

FACS T Cell Killing Data on Panel of Target Positive (CD79b+ and CD20+)and Target Negative (CD79B− and CD20−) Cell Lines

Functional activity of the CD79b×CD3 bispecific and CD79b×CD20×CD3trispecific constructs was assessed at 72 hr time point in an in vitro Tcell killing assay by flow cytometry using cell lines that werevalidated by flow cytometry to have different endogenous expressionlevels of CD79b and CD20 on the cell surface, shown in Table 43.

TABLE 43 CD79b and CD20 Antigen Density of B Lymphoma Cell Lines CD20Antigen CD79b Antigen Density Density (Antigen (Antigen Cell Line CellType Number/cell) Number/cell) HBL-1 Diffuse large B-cell 429,649 73,467lymphoma line OCI-LY10 Diffuse large B-cell 38,885 67,352 lymphoma lineCARNAVAL Diffuse large B-cell 98,176 118,789 lymphoma line K562 Chronicmyelogenous 0 0 leukemia HEL Erythroleukemia 0 0

Target cancer cells were maintained in complete RPMI 1640 (ThermoFisher,catalog #11875093) media containing 10% heat inactivated fetal bovineserum. Prior to the assay, antibodies were made at 3-fold serialdilutions in RPMI 1640 media containing 10% heat inactivated fetalbovine serum, at 4-fold expected final concentration. A volume of 50 μLof medium-diluted bispecific or trispecific Ab in each well of a 96-wellplate were further diluted into 200 μL by adding a mix of target andeffector cell suspension. The target cell lines were harvested bycentrifuge at 400×g for 5 min, washed one time with RPMI 1640 media.Target cancer cells were stain targets with CellTrace CFSE(ThermoFisher; Cat #: C34554) diluted 1/5000. Healthy donor T cells(isolated by CD3—negative selection provided by Discovery Life Sciences)were thawed in complete media (RPMI 1640 media containing 10% heatinactivated fetal bovine serum), counted and suspended in fresh completephenol red-free RPMI 1640 media at 1×10⁶ cells/mL. Target cells and Tcells were mixed to obtain 5:1 effector to target cell ratio. Cellsuspension was added to antibody dilution wells according to platelayout (150 μL/well). Cells were incubated for 72 hours with CD79b×CD3or CD79b×CD20x×CD3 test molecules (100 nM starting concentration at 1:3serial dilutions) at 37° C. All cells were washed with BD stain buffer(BD Biosciences; Cat #554657), centrifuged at 1200 RPM for 3 minutes,with supernatant discarded. Cells were stained for 15 minutes at roomtemperature with Fixable Live/Dead stain (ThermoFisher; Cat #65-0865-14)at a 1:1000 dilution. All cells were washed with BD stain buffer (BDBiosciences; Cat #554657), centrifuged at 1200 RPM for 3 mins, withsupernatant discarded. Cells were then stained for 30 minutes at 4° C.with BD stain buffer containing flow panel antibodies (Table 44),antibodies amount added as listed in the table. All cells were washedwith BD stain buffer (BD Biosciences; Cat #554657), centrifuged at 1200RPM for 3 mins, with supernatant discarded. Cells were analyzed usingFACS Lyric (BD) flow cytometer and percent of cancer cell killing wasgenerated using Cytobank. Percent of cancer cell killing was graphed andIC50 values generated using GraphPad PRISM v.8. Dose response curveswere generated by transforming the x axis values using the formulax=lox. Data was then graphed using non-linear regression curve fitanalysis “log(inhibitor) vs. response-variable slope (four parameter)”.

TABLE 44 Flow Panel Antibodies for T cell killing Assay Amount AntibodyConjugated Catalog LOT added per Name Fluorophore Vendor Number Number:well (μl) CD4 V500 BD Biosciences 560768 9340575  2μ/well CD8 PerCPCy5.5BD Biosciences 560662 9290508 2 μl/well CD69 PE BD Biosciences 5609689049603 10 μl/well  CD25 BV421 BD Biosciences 562443 10302 2 μl/well

CD79b×CD20×CD3 trispecific mediated more potent cytotoxicity as comparedto bispecific constructs in CD79b- and CD20-target positive cell lines.IC50 (pM) values are listed in Table 45. No killing has been observed intarget negative cell lines (FIG. 12A-12B).

TABLE 45 Killing of target positive (CARNAVAL, OCI-Ly10) cell lines(FACS). CARNAVAL HBL-1 OCI-LY10 Protein ID CD79b CD20 CD3 IC50 (nM)*IC50 (nM) IC50 (nM)** 79C3B646 CD9B330 NA CD3B2030 0.29 0.73 >100 nMC923B74 CD9B330 C20B22 CD3B2030 0.35 2.42 24.19 79C3B601 CD9B374 NACD3B2030 NA 2.86 >100 nM C923B38 CD9B374 C20B22 CD3B2030 0.33 2.71 48.5979C3B651 CD9B643 NA CD3B2030 0.25 2.20 >100 nM C923B99 CD9B643 C20B22CD3B2030 0.17 1.68 16.95 *average values of T cell mediated killing from3 independent T cell donors **average values of T cell mediated killingfrom 4 independent T cell donors

Bispecific CD79b×CD3 Mediated Cytotoxicity Against Autologous B-Cells

Functional activity of the CD79b×CD3 bispecific constructs was assessedin an in vitro autologous B cell depletion assay. This functional assayutilizes PBMCs to focus on the killing of primary B cells as well as Tcell activation on donor matched primary cells. Cryo-preserved PBMCsfrom 3 different human donors were incubated for 72 hours with CD79b×CD3test molecules 79C3B646, 79C3B651, and 79C3B601 (300 nM startingconcentration at 1:3 serial dilutions) at 37° C. All cells were washedwith BD stain buffer (BD Biosciences; Cat #554657), centrifuged at 1200RPM for 3 minutes, with supernatant discarded. Cells were stained for 10minutes at room temperature with BD stain buffer containing Fc blockingagent (Accurate Chemical and Scientific Corp; Cat #NB309) and Near IRFixable Live/Dead stain (Invitrogen; Cat #L10119) at a 1:400 dilution.All cells were washed with BD stain buffer (BD Biosciences; Cat#554657), centrifuged at 1200 RPM for 3 mins, with supernatantdiscarded. Cells were then stained for 30 minutes at 4° C. with BD stainbuffer containing flow panel antibodies (Table 46) at a 1:100 dilution.All cells were washed with BD stain buffer (BD Biosciences; Cat#554657), centrifuged at 1200 RPM for 3 mins, with supernatantdiscarded. Cells were analyzed using Intellicyt (Sartorius) flowcytometer and mean fluorescent intensity (MFI) was generated usingForcyt software (Sartorius). MFI was graphed and EC50 values generatedusing GraphPad PRISM v.8. Dose response curves were generated bytransforming the x axis values using the formula x=lox. Data was thengraphed using non-linear regression curve fit analysis “log(agonist) vs.response-variable slope (four parameter)”.

TABLE 46 Flow Panel Antibodies for Autologous B Cell Depletion AssayConjugated Catalog Antibody Name Fluorophore Vendor Number Anti-humanCD25 BV650 BD Biosciences 563719 Anti-Human CD4 BV510 Biolegend 317444Anti-Human CD8 PE-Cy7 Biolegend 301012 Anti-Human CD20 PE Biolegend302306 Anti-Human CD11c AF647 BD Biosciences 565911 Anti-Human CD2 BV605BD Biosciences 740391

CD79b×CD3 bispecific constructs showed a maximum drug mediatedcytotoxicity of 20 percent with low levels of CD4⁺ and CD8⁺ T cellactivation as demonstrated by CD25 expression on these T cell subsets,as shown in FIGS. 13A-13C. The CD79b×CD20×CD3 trispecific has asynergistic effect on drug mediated cytotoxicity when compared tocontrol molecules as shown in Table 47.

TABLE 47 CD79b × CD20 × CD3 EC50 Values and Maximum Cytotoxicity Donor 1Donor 2 Donor 3 EC50 EC50 EC50 Construe Name CD79b Arm CD20 Arm CD3 Arm(nM) C_(max) (nM) C_(max) (nM) C_(max) C923B74 B330-Fab C20B22 scFvCD3B2030 78 34% 23 66% UD* 40% 79C3B646 N/A CD3B2030 40  9%  6 14%  835% C23B98 N/A C20B22 scFv CD3B2030 UD* 11% UD*  5% 88 52% C923B99B643-Fab C20B22 srFv CD3B2030 UD* 35%  9 67% 23 66% 79C3B651 N/ACD3B2030 UD*  5% UD* 13% 17 34% C23B98 N/A C20B22 CD3B2030 UD* 11% UD* 5% 88 52% C923B38 B374-Fab C20B22 scFv CD3B2030  1 24% 50 30% UD* 50%79C3B601 N/A CD3B2030 UD* 10% UD* 10% 20 32% C23B98 N/A C20B22 scFvCD3B2030 UD* 11% UD*  9% 88 52% *Undetermined

Example 6: Biophysical Characterization

Binding Affinity by SPR

General Protocol for SPR Affinity Assessment: Affinity assessment of thebispecific and trispecific constructs against human CD79b were measuredusing recombinantly expressed extracellular domain of CD79b short andlong isoforms (CD9W7.001 and CD9W8.001, respectively) by Surface PlasmonResonance (SPR) using a Biacore 8 k SPR system (Biacore) at 25° C. inHBSP+ buffer. Cross-reactivity of the same antibody panel was alsoassessed against cyno and mouse antigens (CD9W1.001 and CD9W105.001,respectively). Briefly, a C1 sensor chip was immobilized with anti-humanFc (target immobilization levels of >400 RU) using vendor recommendedamino coupling protocol. The test antibodies were captured throughimmobilized anti-Fc and was followed by the injection of different CD79bconstructs at different concentration series (human CD79b short and longisoforms: 30 nM-0.37 nM at 3-fold dilutions; cyno and mouse CD79b: 3000nM-37 nM at 3-fold dilutions). The association and dissociation phaseswere measured for 2 or 3 minutes and 30 minutes, respectively. Bindingof the trispecifics (C923B168 and C923B169) to CD3 was tested byinjecting CD3W220.001 at 100 nM-1.23 nM at 3-fold dilutions, withassociation and dissociation phases were measured for 3 min and 15 min,respectively (CD79b-00478).

The raw binding sensorgrams were processed using Biacore Insightsoftware (Biacore) by double-referencing and the processed sensorgramswere analyzed for cross-reactivity and fitted to a 1:1 Langmuir model toobtain on-rates, off-rates and affinities.

SPR Binding Results: As shown in Table 48 and Table 49, 5he bispecificand trispecific antibodies bound to the human CD79b long isoform (huCD79b long) with affinities from 0.02-0.06 nM, and to the CD79b shortisoform (hu CD79b short) with affinities between 0.27-0.64 nM. Theantibody panel showed very poor cross-reactivity to cyno CD79b (KDestimated >3000 nM) or did not bind to mouse CD79b. C923B168 bindsrecombinant CD3 antigen with an affinity of 0.5 nM. No quantitativekinetics/affinities were reported for those with complex kinetic bindingprofiles using the specified antigens, as noted in the summary tablesbelow.

TABLE 48 Binding affinities for bispecific antibody constructs KD to huKD to hu CD79b CD79b KD to hu Name long (M) short (M) CD3 (M) 79C3B6014.6E−11 5.6E−10 n.d** 79C3B646 2.2E−11 5.8E−10 n.d** 79C3B651 5.2E−113.5E−10 n.d** 79C3B605 n.d* n.d* n.d** 79C3B645 n.d* n.d* n.d** 79C3B650n.d* n.d* n.d** *samples not submittedfor SPR binding analysis**Affinities for CD3 not determined due to complex SPR binding profilesobserved for Cris7b derived CD3 antibodies (historically observedresults).

TABLE 49 Binding affinities for trispecific antibody constructs KD to huKD to hu KD to hu KD to CD79b CD79b CD20 hu CD3 Name long (M) short (M)(M) (M) C923B38  6.5E−11 6.4E−10 n.d** n.d** C923B74  2.3E−11 3.9E−10n.d** n.d** C923B99  4.0E−11 2.7E−10 n.d** n.d** C923B36 n.d* n.d* n.d**n.d** C923B73 n.d* n.d* n.d** n.d** C923B95 n.d* n.d* n.d** n.d**C923B168 1.92E−10 n.d* n.d** 4.96E−10 C923B169 1.64E−10 n.d* n.d** n.d***samples not submittedfor SPR binding analysis **Affinities for CD20 orCD3 not determined due to SPR constraints with CD20 nanodiscs or complexbinding profiles observedfor Cris7b derived CD3 antibodies (historicallyobserved results)

Binding Epitope by HDX-MS

The CD79b epitopes bound by trispecific molecules CD9B374 and CD9B643were mapped by Hydrogen Deuterium Exchange Mass Spectrometry (HDX-MS)according to the following protocol.

General Procedure for HDX-MS Data Acquisition. HDX-MS sample preparationwas performed with automated HDx system (LEAP Technologies, Morrisville,N.C.). The columns and pump were: protease, protease type XIII (proteasefrom Aspergillus saitoi, type XIII)/pepsin column (w/w, 1:1; 2.1×30 mm)(NovaBioAssays Inc., Woburn, Mass.); trap, ACQUITY UPLC BEH C18 VanGuardPre-column (2.1×5 mm) (Waters, Milford, Mass.), analytical, Accucore C18(2.1×100 mm) (Thermo Fisher Scientific, Waltham, Mass.); and LC pump,VH-P10-A (Thermo Fisher Scientific). The loading pump (from the proteasecolumn to the trap column) was set at 600 μL/min with 0.1% aqueousformic acid. The gradient pump (from the trap column to the analyticalcolumn) was set from 9% to 35% acetonitrile in 0.1% aqueous formic acidin 20 min at 100 μL/min.

MS Data Acquisition. Mass spectrometric analyses were carried out usingan LTQ™ Orbitrap Fusion Lumos mass spectrometer (Thermo FisherScientific) with the capillary temperature at 275° C., resolution120,000, and mass range (m/z) 300-1,800.

HDX-MS Data Extraction. BioPharma Finder 3.0 (Thermo Fisher Scientific)was used for the peptide identification of non-deuterated samples priorto the HDX experiments. HDExaminer version 2.5 (Sierra Analytics,Modesto, Calif.) was used to extract centroid values from the MS rawdata files for the HDX experiments.

HDX-MS Data Analysis. The extracted HDX-MS data were further analyzed inExcel. All exchange time points (at pH 6.4 or pH 7.4 at 3.2° C.) wereconverted to the equivalent time points at pH 7.4 and 23° C.

Results

HDX-MS analysis of CD9B374 and CD9B643 indicate binding to a nearlyidentical, conformational epitope of CD79 made up of residues 30-42(SEDRYRNPKGSAC; SEQ ID NO: 253), 50-52 (PRF), 81-86 (EMENP; SEQ ID NO:254), and 144-148 (GFSTL; SEQ ID NO: 255). The residue numbers are thoseof CD79B_Human (P40259).

Thermal Stability of Trispecific CD79b×CD20×CD3 Antibodies by DSC andDSF

The thermal stability of C923B168 and C923B169 was determined byDifferential Scanning calorimetry (DSC) and differential scanningfluorimetry (DSF).

In this characterization, Tonset and Tagg were determined by DSF and theother thermal stability transitions of Tms were determined by DSC. Asshown in Table 50, C923B168 and C923B169 have good thermal stabilitywith Tonset >61° C. and Tm1 >65° C.

TABLE 50 Transition temperatures for trispecific CD79b × CD20 × CD3antibodies: Tonset Tm1 Tm2 Tm3 Tagg Sample ID ° C. σ ° C. σ ° C. σ ° C.σ ° C. σ C923B168.008 61.3 0.15 65.5 0.03 73.5 0.18 77.3 0.07 73.8 0.37C923B169.008 61.7 0.07 68.4 0.03 75.1 0.44 77.7 0.21 74.2 0.5

Example 7: Functional Characterization of CD79×CD20×CD3 TrispecificAntibodies

Binding of Trispecific CD79b×CD20×CD3 Antibodies to Pan T-Cells

Binding of the CD3 arm of CD79b×CD20×CD3 trispecific constructs wasassessed using cryo-preserved, negatively selected, primary human CD3⁺pan T cells. Primary human CD3⁺ pan T cells from three different donorswere incubated for 1 hour with CD79b×CD20×CD3 test molecules C923B169and C923B168 (1 uM starting concentration at 1:3 serial dilutions) at37° C. All cells were washed with BD stain buffer (BD Biosciences; Cat#554657), centrifuged at 1200 RPM for 3 mins, with supernatantdiscarded. Cells were then stained for 20 minutes at 4° C. with BD stainbuffer containing AlexaFluor 647 labeled anti-human IgG secondaryantibody (Jackson Immuno; Cat #109-606-098) at a 1:300 dilution. Allcells were washed with BD stain buffer (BD Biosciences; Cat #554657),centrifuged at 1200 RPM for 3 mins, with supernatant discarded. Cellswere resuspended in 50ul of FACS buffer containing a 1:1000 dilution ofCytox Green viability dye (Invitrogen, Cat #S34860). Cells were analyzedusing Intellicyt (Sartorius) flow cytometer and mean fluorescentintensity (MFI) was generated using Forcyt software (Sartorius). MFI wasgraphed using GraphPad PRISM v.8. Dose response curves were generated bytransforming the x axis values using the formula x=lox. Data was thengraphed using non-linear regression curve fit analysis “log(agonist) vs.response-variable slope (four parameter)”.

All CD79b×CD20×CD3 molecules showed binding on all donor Pan T cellsexpressing endogenous CD3 on the cell surface, shown in Table 51.

TABLE 51 C923B169 and C923B168 CD79b × CD20 × CD3 binding to Pan CD3 Tcells. Pan T cell binding, Pan T cell max binding, Construct EC50 (nM)MFI (×10⁶) Name Donor 1 Donor 2 Donor 3 Donor 1 Donor 2 Donor 3 C923B168125 91 97 1.3 1.0 1.0 C923B169 UD* UD* UD* 0.1 0.03 0.04 UD* =undetermined

FACS T Cell Killing Data on Panel of Target Positive (CD79b⁺ and CD20⁺)Cell Lines

Functional activity of the CD79b×CD20×CD3 trispecific constructs wasassessed at 48 and 72 hr time point in an in vitro T cell killing assayby flow cytometry using cell lines that were validated by flow cytometryto have different endogenous expression levels of CD79b and CD20 on thecell surface, shown in Table 52.

TABLE 52 CD79b and CD20 Antigen Density of B Lymphoma Cell Lines CD20Antigen CD79b Antigen Density Density (Antigen (Antigen Cell Line CellType Number/cell) Number/cell) OCI-LY10 Diffuse large B-cell 38,88567,352 lymphoma line CARNAVAL Diffuse large B-cell 98,176 118,789lymphoma line JEKO-1 Mantle cell lymphoma 280,000 50,000

Target cancer cells were maintained in complete RPMI-1640 (ThermoFisher,catalog #11875093) media containing 10% heat inactivated fetal bovineserum. Prior to the assay, antibodies were made at 3-fold serialdilutions in RPMI 1640 media containing 10% heat inactivated fetalbovine serum, at 4-fold expected final concentration. A volume of 50 μLof medium-diluted bispecific or trispecific Ab in each well of a 96-wellplate were further diluted into 200 μL by adding a mix of target andeffector cell suspension. The target cell lines were harvested bycentrifuge at 400×g for 5 min, washed one time with RPMI 1640 media.Target cancer cells were stain targets with CellTrace CFSE(ThermoFisher; Cat #: C34554) diluted 1/5000. Healthy donor T cells(isolated by CD3—negative selection provided by Discovery Life Sciences)were thawed in complete media (RPMI 1640 media containing 10% heatinactivated fetal bovine serum), counted and suspended in fresh completephenol red-free RPMI 1640 media at 1×10⁶ cells/mL. Target cells and Tcells were mixed to obtain 5:1 effector to target cell ratio. Cellsuspension was added to antibody dilution wells according to platelayout (150 μL/well). Cells were incubated for 48 and 72 hours withCD79b×CD20x×CD3 test molecules C923B169 and C923B168 (100 nM startingconcentration at 1:3 serial dilutions) at 37° C. All cells were washedwith BD stain buffer (BD Biosciences; Cat #554657), centrifuged at 1200RPM for 3 minutes, with supernatant discarded. Cells were stained for 15minutes at room temperature with Fixable Live/Dead stain (ThermoFisher;Cat #65-0865-14) at a 1:1000 dilution. All cells were washed with BDstain buffer (BD Biosciences; Cat #554657), centrifuged at 1200 RPM for3 mins, with supernatant discarded. Cells were then stained for 30minutes at 4° C. with BD stain buffer containing flow panel antibodies(Table 53), antibodies amount added as listed in the table. All cellswere washed with BD stain buffer (BD Biosciences; Cat #554657),centrifuged at 1200 RPM for 3 mins, with supernatant discarded. Cellswere analyzed using FACS Lyric (BD) flow cytometer and percent of cancercell killing was generated using Cytobank. Percent of cancer cellkilling was graphed and IC50 values generated using GraphPad PRISM v.8.Dose response curves were generated by transforming the x axis valuesusing the formula x=lox. Data was then graphed using non-linearregression curve fit analysis “log(inhibitor) vs. response-variableslope (four parameter)”.

TABLE 53 Flow Panel Antibodies for T cell killing Assay Amount AntibodyConjugated Catalog LOT added per Name Fluorophore Vendor Number Number:well (μl) CD4 V500 BD 560768 9340575  2/well Biosciences CD8 PerCPCy5.5BD 560662 9290508 2 ul/well Biosciences CD69 PE BD 560968 9049603 10ul/well  Biosciences CD25 BV421 BD 562443 10302 2 ul/well Biosciences

CD79b×CD20×CD3 trispecific mediated potent cytotoxicity. IC50 (nM)values and Max killing values are listed in Table 54 and Table 55.

TABLE 54 C923B169 and C923B168 CD79b × CD20 × CD3 killing of targetpositive (CARNAVAL, OCI-Ly10, JEK0-1) cell lines (FACS) at 48 hours.IC50 (nM) and percent of maximal killing are listed in the table.Average values from 2 independent T cell donors. Killing Killing KillingKilling Killing Killing CARNAVAL CARNAVAL JEKO-1 JEKO-1 OCI-Ly10OCI-Ly10 1:1 48 hr 5:1 48 hr 1:1 48 hr 5:1 48 hr 1:1 48 hr 5:14 8 hrProtein ID IC50 Max IC50 Max IC50 Max IC50 Max IC50 Max IC50 MaxC923B169 110.3 49.7% 0.179 87.2% 27.307 60.8% 0.027 93.5% >10022.2% >100 19.7% C923B168 13.6 58.8% 0.012 95.0% 7.466 65.3% 0.00296.7% >100 25.1% >100 29.8%

TABLE 55 C923B169 and C923B168 CD79b × CD20 × CD3 killing of targetpositive CARNAVAL, OCI-Ly10, JEK0-1) cell lines (FACS) at 72 hours. IC50(nM) and percent of maximal killing are listed in the table. Averagevalues from 2 independent T cell donors. Killing Killing Killing KillingKilling Killing CARNAVAL; CARNAVAL; JEKO-1; JEKO-1; OCI-Ly10; OCI-Ly10;1:1 72 hr 5:1 72 hr 1:1 72 hr 5:1 72 hr 1:1 72 hr 5:1 72 hr Protein IDIC50 Max IC50 Max IC50 Max IC50 Max IC50 Max IC50 Max C923B169 50.1566.8% 0.026 98.3% 0.087 82.2 0.003 99.4% >100 43.5% 24.58 70.0% C923B16815.53 75.9% 0.003 99.3% 0.010 85.8% 0.001 99.6% 0.81 61.0% 0.51 90.7%

C923B169 and C923B168 CD79b×CD20×CD3 Mediated Cytotoxicity AgainstAutologous B-Cells

Functional activity of the C923B169 and C923B168 CD79b×CD20×CD3constructs was assessed in an in vitro autologous B cell depletionassay. This functional assay utilizes PBMCs to focus on the killing ofprimary B cells as well as T cell activation on donor matched primarycells. Cryo-preserved PBMCs from 3 different human donors were incubatedfor 72 hours with CD79b×CD20×CD3 test molecules C923B169 and C923B168(300 nM starting concentration at 1:3 serial dilutions) at 37° C. Allcells were washed with BD stain buffer (BD Biosciences; Cat #554657),centrifuged at 1200 RPM for 3 minutes, with supernatant discarded. Cellswere stained for 10 minutes at room temperature with BD stain buffercontaining Fc blocking agent (Accurate Chemical and Scientific Corp; Cat#NB309) and Near IR Fixable Live/Dead stain (Invitrogen; Cat #L10119) ata 1:400 dilution. All cells were washed with BD stain buffer (BDBiosciences; Cat #554657), centrifuged at 1200 RPM for 3 mins, withsupernatant discarded. Cells were then stained for 30 minutes at 4° C.with BD stain buffer containing flow panel antibodies (Table 56) at a1:100 dilution. All cells were washed with BD stain buffer (BDBiosciences; Cat #554657), centrifuged at 1200 RPM for 3 mins, withsupernatant discarded. Cells were analyzed using Intellicyt (Sartorius)flow cytometer. EC50 values generated using GraphPad PRISM v.8. Doseresponse curves were generated by transforming the x axis values usingthe formula x=lox. Data was then graphed using non-linear regressioncurve fit analysis “log(agonist) vs. response-variable slope (fourparameter)”.

TABLE 56 Flow Panel Antibodies for Autologous B Cell Depletion AssayConjugated Catalog Antibody Name Fluorophore Vendor Number Anti-humanCD25 BV650 BD Biosciences 563719 Anti-Human CD4 BV510 Biolegend 317444Anti-Human CD8 PE-Cy7 Biolegend 301012 Anti-Human CD20 PE Biolegend302306 Anti-Human CD11c AF647 BD Biosciences 565911 Anti-Human CD2 BV605BD Biosciences 740391

CD79b×CD20×CD3 C923B169 and C923B168 constructs showed a maximum drugmediated cytotoxicity of 69-95 percent (Table 57) with low levels ofCD4⁺ and CD8⁺ T cell activation as demonstrated by CD25 expression onthese T cell subsets.

TABLE 57 C923B169 and C923B168 CD79b × CD20 × CD3 killing of B cell inthe primary autologous B cell depletion assay at 72 hours. EC50 (nM) andpercent of maximal killing are listed in the table. Values from 3independent T cell donors listed. D329465 D198013 D221837 Average valuesEC50 Max Kill EC50 Max Kill EC50 Max Kill EC50 Max Kill Name (nM) (%)(nM) (%) (nM) (%) (nM) (%) C923B168 0.1 69 0.02 92 0.1 95 0.07 84C923B169 2.0 69 1.70 92 6.30 81 2.80 80

Example 8: Generation of Bispecific PSMAxCD3 Antibodies Example 8.1:FAB-ARM Exchange of Anti-PSMA and Anti-CD3 Antibodies

The formation of bispecific antibodies requires two parental monoclonalantibodies (mAbs), one specific for the targeting arm (e.g. PMSA) andone specific for the effector arm (e.g. CD3). Selected monospecificanti-PSMA and anti-CD3 antibodies were expressed as IgG1/κ engineered tohave L234A, L235A and D265S substitutions for cF silencing, (EUnumbering). Selected monospecific anti-PSMA and anti-CD3 antibodies arealso expressed as IgG4 antibodies. Mutations designed to promoteselective heterodimerization of the Fc domain were also engineered inthe Fc domain.

The monospecific antibodies were expressed in CHO cell lines under CMVpromoters as described above). The parental PSMA and CD3 antibodies werepurified using a protein A column with an elution buffer of 100 mM NaAcpH3.5 and a neutralization puffer of 2 M Tris pH 7.5 and 150 mM NaCl.The anti-PSMA and anti-CD3 monoclonal antibodies were dialyzed intoD-PBS, pH 7.2 buffer.

For DuoBody antibodies, post purification of parental monospecificantibodies, bispecific PSMAxCD3 antibodies were generated by mixing theparental PSMA antibodies with the desired parental CD3 antibody underreducing conditions in 75 mM cysteamine-HCl and incubated overnight atroom temperature for in vitro Fab arm exchange as described in Int.Patent Publ. No. WO2011/131746. The recombination reactions were basedon molar ratios, where a set amount of PSMA antibody (e.g., 10 mg, or˜74.6 nanomoles) was combined with CD3 antibody (e.g., ˜70.87nanomoles), where the PSMA antibody was added in a 5% excess of the CD3antibody. The concentrations of the PSMA antibody stocks varied from 0.8to 6 mg/mL, and the volumes the recombination reactions varied for eachpairing. The recombinations were subsequently dialyzed overnight againstPBS to remove the reductant. The PSMAxCD3 bispecific antibody reactionswere performed with an excess of the PSMA antibody (ratio) to minimizethe amount of unreacted CD3 parental antibody remaining afterrecombination.

Other bispecifics were generated via co-transfection of HC1:HC2:LC2,typically at a DNA ratio of 1:1:3. Purification was performed by proteinA chromatography and CH1 affinity capture, followed by an ionexchange-based chromatography.

Exemplary PSMAxCD3 multispecific antibodies are provided in Tables 58through 63.

TABLE 58 PSMA × CD3 Bispecific Antibodies: Clone Descriptions NameBispecific Description PS3B1353 HC1 (F405L): CD3B376 × HC2 (K409R):PSMB896 PS3B1505 HC1 CD3B376-Fab × HC2 PSMB896-G100A IgG1 DuoBodyPS3B1508 HC1 (Knob3): CD3W245-LH-scFv; HC2 (Hole3-RF)PSMB896-G100A-Fab-RF: IgG1 AAS PS3B1917 HC1 (ZWA w/o K447_RF):CD3B376-Fab, HC2 (ZWB w/o K447): PSMA_P72_A10-HC-G54E-scFv LH PS3B1918HC1 (ZWA w/o K447_RF): CD3B376-Fab, HC2 (ZWB w/o K447):PSMA_P72_D01-HC-D95E-scFv LH PS3B1919 HC1 (ZWA w/o K447_RF):CD3B376-Fab; HC2 (ZWB w/o K447): PSMA_P75_F01, LH PS3B1920 HC1 (ZWA w/oK447_RF): CD3B376-Fab; HC2 (ZWB w/o K447): PSMA_P72_F07, LH PS3B1921 HC1(ZWA w/o K447_RF): CD3B376-Fab; HC2 (ZWB w/o K447): PSMA_P72_E07, LHPS3B1922 HC1 (ZWA w/o K447_RF): CD3B376-Fab; HC2 (ZWB w/o K447):PSMA_P72_D01, LH PS3B1923 HC1 (ZWA w/o K447_RF): CD3B376-Fab; HC2 (ZWBw/o K447): PSMA_P72_C01, LH PS3B1924 HC1 (ZWA w/o K447_RF): CD3B376-Fab;HC2 (ZWB w/o K447): PSMA_P72_A10, LH PS3B1925 HC1 (ZWA w/o K447_RF):CD3B376-Fab; HC2 (ZWB w/o K447): PSMA_P70_F02, LH PS3B1926 HC1 (ZWA w/oK447_RF): CD3B376-Fab; HC2 (ZWB w/o K447): PSMA_P72_G02, HL PS3B1927 HC1(ZWA w/o K447_RF): CD3B376-Fab; HC2 (ZWB w/o K447): PSMA_P72_C01, HLPS3B1928 HC1 (ZWA w/o K447_RF): CD3B376-Fab; HC2 (ZWB w/o K447):PSMA_P72_A11, HL PSMB1041 HC1 (ZWA): B23B62-Fab; HC2 (ZWB):PSMA_P70_F02, LH PSMB1045 HC1 (ZWA): B23B62-Fab; HC2 (ZWB):PSMA_P72_A10, LH PSMB1047 HC1 (ZWA): B23B62-Fab; HC2 (ZWB):PSMA_P72_C01, LH PSMB1049 HC1 (ZWA): B23B62-Fab; HC2 (ZWB):PSMA_P72_D01, LH PSMB1051 HC1 (ZWA): B23B62-Fab; HC2 (ZWB):PSMA_P72_E07, LH PSMB1052 HC1 (ZWA): B23B62-Fab; HC2 (ZWB):PSMA_P72_F07, LH PSMB1060 HC1 (ZWA): B23B62-Fab; HC2 (ZWB):PSMA_P75_F01, LH PSMB1068 HC1 (ZWA): B23B62-Fab; HC2 (ZWB):PSMA_P72_A11, HL PSMB1069 HC1 (ZWA): B23B62-Fab; HC2 (ZWB):PSMA_P72_C01, HL PSMB1075 HC1 (ZWA): B23B62-Fab; HC2 (ZWB):PSMA_P72_G02, HL PSMB2908 HC1 (ZWA w/o K447): B23B62-Fab, HC2 (ZWB w/oK447): PSMA_P72_D01-HC-D95E-scFv LH PSMB2909 HC1 (ZWA w/o K447):B23B62-Fab, HC2 (ZWB w/o K447)): PSMA_P72_A10-HC-G54E-scFv LH PS3B1391HC 1: N-term scFv LH CD3B2030 N106A LH-scFv (MA), HC 2:N-term_Fab_PSMHB49SC1133_011A11_1 PS3B1396 HC1 (Knob3):CD3B2030-N106A-scFv LH, HC2 (Hole3): PSMB896-G100A-Fab

TABLE 59 PSMA x CD3 Bispecific Antibodies: CD3 Arm Descriptions SEQ SEQCD3 Arm Name HC1 ID NO. LC1 ID NO. Description PS3B1353QVQLQQSGPRLVRPSQTLSLTC 1188 QSALTQPASVSG 1189 withAISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376 RGLEWLGRTYYRSKWLYDYAVSSSNIGTYKFVSW arm VKSRITVNPDTSRNQFTLQL YQQHPDKAPKV (CD3B891NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477 YWGQGTLVTVSSASTKGPSVFPSSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDY ASLTISGLQAED CD3B2197FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 in AVLQSSGLYSLSSVVTVPSSSLGSGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKV TVLGQPKAAPSVEPKSCDKTHTCPPCPAPEAAGG TLFPPSSEELQAN PSVFLFPPKPKDTLMISRTPEVTKATLVCLISDFY CVVVSVSHEDPEVKFNWYVDG PGAVTVAWKAD VEVHNAKTKPREEQYNSTYRVSSPVKAGVETTT VSVLTVLHQDWLNGKEYKCK PSKQSNNKYAAS VSNKALPAPIEKTISKAKGQPRESYLSLTPEQWKS PQVYTLPPSREEMTKNQVSLTC HRSYSCQVTHEG LVKGFYPSDIAVEWESNGQPENSTVEKTVAPTEC NYKTTPPVLDSDGSFLLYSKLT S VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK PS3B1505 QVQLQQSGPRLVRPSQTLSLTC 1190 QSALTQPASVSG 1191with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376 RGLEWLGRTYYRSKWLYDYASSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV (CD3B891NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477 YWGQGTLVTVSSASTKGPSVFPSSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDY ASLTISGLQAED CD3B2197FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 in AVLQSSGLYSLSSVVTVPSSSLGSGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKV TVLGQPKAAPSVEPKSCDKTHTCPPCPAPEAAGG TLFPPSSEELQAN PSVFLFPPKPKDTLMISRTPEVTKATLVCLISDFY CVVVSVSHEDPEVKFNWYVDG PGAVTVAWKAD VEVHNAKTKPREEQYNSTYRVSSPVKAGVETTT VSVLTVLHQDWLNGKEYKCK PSKQSNNKYAAS VSNKALPAPIEKTISKAKGQPRESYLSLTPEQWKS PQVYTLPPSREEMTKNQVSLTC HRSYSCQVTHEG LVKGFYPSDIAVEWESNGQPENSTVEKTVAPTEC NYKTTPPVLDSDGSFLLYSKLT S VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK PS3B1508 DIQMTQSPSSLSASVGDRVTITC 1192 NA 1193 withRARQSIGTAIHWYQQKPGKAP CD3W245 KLLIKYASESISGVPSRFSGSGS armGTDFTLTISSLQPEDFATYYCQQ (CD3B2183 SGSWPYTFGQGTKLEIKGGSEG withoutKSSGSGSESKSTGGSEVQLVES K477) GGGLVKPGGSLRLSCAASGFTF SRYNMNWVRQAPGKGLEWVSSISTSSNYIYYADSVKGRFTFSR DNAKNSLDLQMSGLRAEDTAI YYCTRGWGPFDYWGQGTLVTVSSEPKSSDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPE VTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKC KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSL WCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGK PS3B1917 QVQLQQSGPRLVRPSQTLSLTC1194 QSALTQPASVSG 1195 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1918 QVQLQQSGPRLVRPSQTLSLTC1196 QSALTQPASVSG 1197 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1919 QVQLQQSGPRLVRPSQTLSLTC1198 QSALTQPASVSG 1199 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1920 QVQLQQSGPRLVRPSQTLSLTC1200 QSALTQPASVSG 1201 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1921 QVQLQQSGPRLVRPSQTLSLTC1202 QSALTQPASVSG 1203 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1922 QVQLQQSGPRLVRPSQTLSLTC1204 QSALTQPASVSG 1205 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1923 QVQLQQSGPRLVRPSQTLSLTC1206 QSALTQPASVSG 1207 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1924 QVQLQQSGPRLVRPSQTLSLTC1208 QSALTQPASVSG 1209 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1925 QVQLQQSGPRLVRPSQTLSLTC1210 QSALTQPASVSG 1211 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1926 QVQLQQSGPRLVRPSQTLSLTC1212 QSALTQPASVSG 1213 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1927 QVQLQQSGPRLVRPSQTLSLTC1214 QSALTQPASVSG 1215 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PS3B1928 QVQLQQSGPRLVRPSQTLSLTC1216 QSALTQPASVSG 1217 with AISGDSVFNNNAAWSWIRQSPS SPGQSITISCTGT CD3B376RGLEWLGRTYYRSKWLYDYA SSNIGTYKFVSW arm VSVKSRITVNPDTSRNQFTLQL YQQHPDKAPKV(CD3B891 NSVTPEDTALYYCARGYSSSFD LLYEVSKRPSGV without K477YWGQGTLVTVSSASTKGPSVFP SSRFSGSKSGNT or LAPSSKSTSGGTAALGCLVKDYASLTISGLQAED CD3B2197 FPEPVTVSWNSGALTSGVHTFP QADYHCVSYAG with K477 inAVLQSSGLYSLSSVVTVPSSSLG SGTLLFGGGTKL HC1) TQTYICNVNHKPSNTKVDKKVTVLRTVAAPSVF EPKSCDKTHTCPPCPAPEAAGG IFPPSDEQLKSGTPSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYP CVVVSVSHEDPEVKFNWYVDG REAKVQWKVDNVEVHNAKTKPREEQYNSTYRV ALQSGNSQESVT VSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLSVSNKALPAPIEKTISKAKGQPRE STLTLSKADYEK PQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQLVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNR NYKTTPPVLDSDGSFALVSKLT GECVDKSRWQQGNVFSCSVMHEAL HNRFTQKSLSLSPG PSMB1041 QITLKESGPTLVKPTQTLTLTCT1218 DIVMTQSPDSLA 1219 Null CD3 FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR armALEWLAHIYWDDDKRYNPSLK ASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQPDPVDTATYYCARLYGFTYGFA PKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSGLAPSSKSTSGGTAALGCLVKDY TDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPELLGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVDVSHEDPEVKFNWYVD REAKVQWKVDN GVEVHNAKTKPREEQYNSTYR ALQSGNSQESVTVVSVLTVLHQDWLNGKEYKC EQDSKDSTYSLS KVSNKALPAPIEKTISKAKGQP STLTLSKADYEKREPQVYVYPPSREEMTKNQVSL HKVYACEVTHQ TCLVKGFYPSDIAVEWESNGQP GLSSPVTKSFNRENNYKTTPPVLDSDGSFALVSK GEC LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGPSMB1045 QITLKESGPTLVKPTQTLTLTCT 1220 DIVMTQSPDSLA 1221 Null CD3FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPELLGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVDVSHEDPEVKFNWYVD REAKVQWKVDN GVEVHNAKTKPREEQYNSTYR ALQSGNSQESVTVVSVLTVLHQDWLNGKEYKC EQDSKDSTYSLS KVSNKALPAPIEKTISKAKGQP STLTLSKADYEKREPQVYVYPPSREEMTKNQVSL HKVYACEVTHQ TCLVKGFYPSDIAVEWESNGQP GLSSPVTKSFNRENNYKTTPPVLDSDGSFALVSK GEC LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGPSMB1047 QITLKESGPTLVKPTQTLTLTCT 1222 DIVMTQSPDSLA 1223 Null CD3FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPELLGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVDVSHEDPEVKFNWYVD REAKVQWKVDN GVEVHNAKTKPREEQYNSTYR ALQSGNSQESVTVVSVLTVLHQDWLNGKEYKC EQDSKDSTYSLS KVSNKALPAPIEKTISKAKGQP STLTLSKADYEKREPQVYVYPPSREEMTKNQVSL HKVYACEVTHQ TCLVKGFYPSDIAVEWESNGQP GLSSPVTKSFNRENNYKTTPPVLDSDGSFALVSK GEC LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGPSMB1049 QITLKESGPTLVKPTQTLTLTCT 1224 DIVMTQSPDSLA 1225 Null CD3FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPELLGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVDVSHEDPEVKFNWYVD REAKVQWKVDN GVEVHNAKTKPREEQYNSTYR ALQSGNSQESVTVVSVLTVLHQDWLNGKEYKC EQDSKDSTYSLS KVSNKALPAPIEKTISKAKGQP STLTLSKADYEKREPQVYVYPPSREEMTKNQVSL HKVYACEVTHQ TCLVKGFYPSDIAVEWESNGQP GLSSPVTKSFNRENNYKTTPPVLDSDGSFALVSK GEC LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGPSMB1051 QITLKESGPTLVKPTQTLTLTCT 1226 DIVMTQSPDSLA 1227 Null CD3FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPELLGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVDVSHEDPEVKFNWYVD REAKVQWKVDN GVEVHNAKTKPREEQYNSTYR ALQSGNSQESVTVVSVLTVLHQDWLNGKEYKC EQDSKDSTYSLS KVSNKALPAPIEKTISKAKGQP STLTLSKADYEKREPQVYVYPPSREEMTKNQVSL HKVYACEVTHQ TCLVKGFYPSDIAVEWESNGQP GLSSPVTKSFNRENNYKTTPPVLDSDGSFALVSK GEC LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGPSMB1052 QITLKESGPTLVKPTQTLTLTCT 1228 DIVMTQSPDSLA 1229 Null CD3FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPELLGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVDVSHEDPEVKFNWYVD REAKVQWKVDN GVEVHNAKTKPREEQYNSTYR ALQSGNSQESVTVVSVLTVLHQDWLNGKEYKC EQDSKDSTYSLS KVSNKALPAPIEKTISKAKGQP STLTLSKADYEKREPQVYVYPPSREEMTKNQVSL HKVYACEVTHQ TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSK GLSSPVTKSFNR LTVDKSRWQQGNVFSCSVMHE GECALHNRFTQKSLSLSPG PSMB1060 QITLKESGPTLVKPTQTLTLTCT 1230 DIVMTQSPDSLA 1231Null CD3 FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPELLGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVDVSHEDPEVKFNWYVD REAKVQWKVDN GVEVHNAKTKPREEQYNSTYR ALQSGNSQESVTVVSVLTVLHQDWLNGKEYKC EQDSKDSTYSLS KVSNKALPAPIEKTISKAKGQP STLTLSKADYEKREPQVYVYPPSREEMTKNQVSL HKVYACEVTHQ TCLVKGFYPSDIAVEWESNGQP GLSSPVTKSFNRENNYKTTPPVLDSDGSFALVSK GEC LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGPSMB1068 QITLKESGPTLVKPTQTLTLTCT 1232 DIVMTQSPDSLA 1233 Null CD3FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPELLGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVDVSHEDPEVKFNWYVD REAKVQWKVDN GVEVHNAKTKPREEQYNSTYR ALQSGNSQESVTVVSVLTVLHQDWLNGKEYKC EQDSKDSTYSLS KVSNKALPAPIEKTISKAKGQP STLTLSKADYEKREPQVYVYPPSREEMTKNQVSL HKVYACEVTHQ TCLVKGFYPSDIAVEWESNGQP GLSSPVTKSFNRENNYKTTPPVLDSDGSFALVSK GEC LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGPSMB1069 QITLKESGPTLVKPTQTLTLTCT 1234 DIVMTQSPDSLA 1235 Null CD3FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPELLGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVDVSHEDPEVKFNWYVD REAKVQWKVDN GVEVHNAKTKPREEQYNSTYR ALQSGNSQESVTVVSVLTVLHQDWLNGKEYKC EQDSKDSTYSLS KVSNKALPAPIEKTISKAKGQP STLTLSKADYEKREPQVYVYPPSREEMTKNQVSL HKVYACEVTHQ TCLVKGFYPSDIAVEWESNGQP GLSSPVTKSFNRENNYKTTPPVLDSDGSFALVSK GEC LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGPSMB1075 QITLKESGPTLVKPTQTLTLTCT 1236 DIVMTQSPDSLA 1237 Null CD3FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPELLGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVDVSHEDPEVKFNWYVD REAKVQWKVDN GVEVHNAKTKPREEQYNSTYR ALQSGNSQESVTVVSVLTVLHQDWLNGKEYKC EQDSKDSTYSLS KVSNKALPAPIEKTISKAKGQP STLTLSKADYEKREPQVYVYPPSREEMTKNQVSL HKVYACEVTHQ TCLVKGFYPSDIAVEWESNGQP GLSSPVTKSFNRENNYKTTPPVLDSDGSFALVSK GEC LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGPSMB2908 QITLKESGPTLVKPTQTLTLTCT 1238 DIVMTQSPDSLA 1239 Null CD3FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPEAAGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVSVSHEDPEVKFNWYVDG REAKVQWKVDN VEVHNAKTKPREEQYNSTYRV ALQSGNSQESVTVSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLS VSNKALPAPIEKTISKAKGQPRE STLTLSKADYEKPQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQ LVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNRNYKTTPPVLDSDGSFALVSKLT GEC VDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG PSMB2909QITLKESGPTLVKPTQTLTLTCT 1240 DIVMTQSPDSLA 1241 Null CD3FSGFSLSTSGMGVSWIRQPPGK VSLGERATINCR arm ALEWLAHIYWDDDKRYNPSLKASQSVDYNGISY SRLTITKDTSKNQVVLTMTNM MHWYQQKPGQP DPVDTATYYCARLYGFTYGFAPKLLIYAASNPES YWGQGTLVTVSSASTKGPSVFP GVPDRFSGSGSG LAPSSKSTSGGTAALGCLVKDYTDFTLTISSLQAE FPEPVTVSWNSGALTSGVHTFP DVAVYYCQQIIEAVLQSSGLYSLSSVVTVPSSSLG DPWTFGQGTKV TQTYICNVNHKPSNTKVDKKV EIKRTVAAPSVFIEPKSCDKTHTCPPCPAPEAAGG FPPSDEQLKSGT PSVFLFPPKPKDTLMISRTPEVT ASVVCLLNNFYPCVVVSVSHEDPEVKFNWYVDG REAKVQWKVDN VEVHNAKTKPREEQYNSTYRV ALQSGNSQESVTVSVLTVLHQDWLNGKEYKCK EQDSKDSTYSLS VSNKALPAPIEKTISKAKGQPRE STLTLSKADYEKPQVYVYPPSREEMTKNQVSLTC HKVYACEVTHQ LVKGFYPSDIAVEWESNGQPEN GLSSPVTKSFNRNYKTTPPVLDSDGSFALVSKLT GEC VDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG PS3B1391EIVLTQSPATLSASPGERVTLSC 1455 NA 1456 CD3B2030 SASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGS GRDYTLTISSLEPEDFAVYYCQ QWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQ SGAEVKKPGSSVKVSCKASGY TFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTL TADKSTSTAYMELSSLRSEDTA VYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPP CPAPEAAGGPSVFLFPPKPKDT LMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPR EEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLWCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSL SLSPGK PS3B1396EIVLTQSPATLSASPGERVTLSC 1457 NA 1458 CD3B2030 SASSSVSYMNWYQQKPGQAPRRWIYDSSKLASGVPARFSGSGS GRDYTLTISSLEPEDFAVYYCQ QWSRNPPTFGGGTKVEIKGGSEGKSSGSGSESKSTGGSQVQLVQ SGAEVKKPGSSVKVSCKASGY TFTRSTMHWVKQAPGQGLEWIGYINPSSAYTNYNQKFQGRVTL TADKSTSTAYMELSSLRSEDTA VYYCASPQVHYDYAGFPYWGQGTLVTVSSEPKSSDKTHTCPP CPAPEAAGGPSVFLFPPKPKDT LMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPR EEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLWCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSL SLSPGK

TABLE 60 PSMA x CD3 Bispecific Antibodies: PSMA Arm Descriptions SEQ SEQPSMA Arm Name HC2 ID NO. LC2 ID NO. Description PS3B1353EVQLVESGGGLVQPGGSLRLSC 1242 QSVLTQPPSVSA 1243 PSMB896AASGFTFSSYAMSWVRQAPGK APGQKVTISCSG GLEWVSAISGGIGSTYYADSVK SSSNIGINYVSWGRFTISRDNSKNTLWLQMNSLR YQQLPGTAPKLL AEDTAVYYCAKDGVGATPYYF IYDNNKRPSGIPDYWGQGTLVTVSSASTKGPSV DRFSGSKSGTSA FPLAPSSKSTSGGTAALGCLVK TLGITGLQTGDEDYFPEPVTVSWNSGALTSGVHT ADYYCGTWDSS FPAVLQSSGLYSLSSVVTVPSSS LSAVVFGGGTKLLGTQTYICNVNHKPSNTKVDK TVLGQPKAAPSV KVEPKSCDKTHTCPPCPAPEAA TLFPPSSEELQANGGPSVFLFPPKPKDTLMISRTPE KATLVCLISDFY VTCVVVSVSHEDPEVKFNWYV PGAVTVAWKADDGVEVHNAKTKPREEQYNSTY SSPVKAGVETTT RVVSVLTVLHQDWLNGKEYKC PSKQSNNKYAASKVSNKALPAPIEKTISKAKGQP SYLSLTPEQWKS REPQVYTLPPSREEMTKNQVSL HRSYSCQVTHEGTCLVKGFYPSDIAVEWESNGQP STVEKTVAPTEC ENNYKTTPPVLDSDGSFFLYSR SLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK PS3B1505 EVQLVESGGGLVQPGGSLRLSC1244 QSVLTQPPSVSA 1245 PSMB896- AASGFTFSSYAMSWVRQAPGK APGQKVTISCSG G100AGLEWVSAISGGIGSTYYADSVK SSSNIGINYVSW GRFTISRDNSKNTLWLQMNSLR YQQLPGTAPKLLAEDTAVYYCAKDAVGATPYYF IYDNNKRPSGIP DYWGQGTLVTVSSASTKGPSV DRFSGSKSGTSAFPLAPSSKSTSGGTAALGCLVK TLGITGLQTGDE DYFPEPVTVSWNSGALTSGVHT ADYYCGTWDSSFPAVLQSSGLYSLSSVVTVPSSS LSAVVFGGGTKL LGTQTYICNVNHKPSNTKVDK TVLGQPKAAPSVKVEPKSCDKTHTCPPCPAPEAA TLFPPSSEELQAN GGPSVFLFPPKPKDTLMISRTPEKATLVCLISDFY VTCVVVSVSHEDPEVKFNWYV PGAVTVAWKAD DGVEVHNAKTKPREEQYNSTYSSPVKAGVETTT RVVSVLTVLHQDWLNGKEYKC PSKQSNNKYAAS KVSNKALPAPIEKTISKAKGQPSYLSLTPEQWKS REPQVYTLPPSREEMTKNQVSL HRSYSCQVTHEG TCLVKGFYPSDIAVEWESNGQPSTVEKTVAPTEC ENNYKTTPPVLDSDGSFFLYSR S LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK PS3B1508 EVQLVESGGGLVQPGGSLRLSC 1246 QSVLTQPPSVSA 1247PSMB896- AASGFTFSSYAMSWVRQAPGK APGQKVTISCSG G100A GLEWVSAISGGIGSTYYADSVKSSSNIGINYVSW GRFTISRDNSKNTLWLQMNSLR YQQLPGTAPKLL AEDTAVYYCAKDAVGATPYYFIYDNNKRPSGIP DYWGQGTLVTVSSASTKGPSV DRFSGSKSGTSA FPLAPSSKSTSGGTAALGCLVKTLGITGLQTGDE DYFPEPVTVSWNSGALTSGVHT ADYYCGTWDSS FPAVLQSSGLYSLSSVVTVPSSSLSAVVFGGGTKL LGTQTYICNVNHKPSNTKVDK TVLGQPKAAPSV KVEPKSCDKTHTCPPCPAPEAATLFPPSSEELQAN GGPSVFLFPPKPKDTLMISRTPE KATLVCLISDFY VTCVVVSVSHEDPEVKFNWYVPGAVTVAWKAD DGVEVHNAKTKPREEQYNSTY SSPVKAGVETTT RVVSVLTVLHQDWLNGKEYKCPSKQSNNKYAAS KVSNKALPAPIEKTISKAKGQP SYLSLTPEQWKS REPQVYTLPPSREEMTKNQVSLHRSYSCQVTHEG SCAVKGFYPSDIAVEWESNGQP STVEKTVAPTEC ENNYKTTPPVLDSDGSFFLVSKS LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGK PS3B1917SYELMQPPSVSVSPGQTARITCS 1248 NA 1249 PSMA_P72 GDALPKQYAYWYQQKPGQAPA10-HC- VLVIYKDSERPSGIPVRFSGSSS G54E GTTVTLTITGVQAEDEADYYCQSADSSGTYVFGTGTKVTVLGGS EGKSSGSGSESKSTGGSQVQLV ESGGGVVQPGRSLRLSCAASGFTFSSYNMNWVRQAPGKGLEW VAIIYYDESNKYYADSVKGRFT ISRDISKNTLYLQMNSLRAEDTAVYYCARERGRDYYGMDVWG QGTTVTVSSEPKSSDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPE VKFNWYVDGVEVHNAKTKPR EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYVLPPSREE MTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDS DGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLS LSPGPS3B1918 QSVLTQPASVSGSPGQSITISCT 1250 NA 1251 PSMA_P_72_GTSSDVGGYNYVSWYQQHPG D01-HC- KAPKLMIYEVSNRPSGVSNRFS D95EGSKSGNTASLTISGLQAEDEAD YYCSSYTSSYTYVFGTGTKLTV LGGSEGKSSGSGSESKSTGGSEVQLVESGGDLVQPGGSLRLSCA ASGFTFNNYNMNWVRQAPGK GLEWVSHISTSSSNKYYADSVKGRFSISRDIAKNSMYLQMNSLR DEDTAVYYCAREGVGADYGD YYYYGMDVWGQGTTVTVSSEPKSSDKTHTCPPCPAPEAAGGP SVFLFPPKPKDTLMISRTPEVTC VVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVS NKALPAPIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLV KGFYPSDIAVEWESNGQPENNY LTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPG PS3B1919 EIVLTQSPGTLSVSPGERATLSC 1252NA 1253 PSMA_P75_ RASQSVRSNLAWYQQKPGQAP F01 RLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCHQ YNDWPPYTFGQGTKLEIKGGSE GKSSGSGSESKSTGGSQVQLQESGGGVVQPGRSLRLSCAASGFT FSTYGMHWVRQAPGKGLEWV AFISYDGSNKYYADSVKGRFTISRDNSKHTLYLQMNSLRAEDT AVYYCAGRDNLRFLEWFMDV WGQGTTVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKD TLMISRTPEVTCVVVSVSHEDP EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPSDI AVEWESNGQPENNYLTWPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPG PS3B1920 SYELTQPPSVSVAPGQTARITCG 1254 NA1255 PSMA_P72_ GNNIGSKSVHWYQQKPGQAPV F07 LVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQV WDSSTDHVVFGGGTKLTVLGG SEGKSSGSGSESKSTGGSEVQLVESGGGVVQPGRSLRLSCAASG FTFSSYGMNWVRQAPGKGLEW VAVTSYDGSNKYYADSVKGRFTISRDISKNTLYLQMSSLRAEDT AVYYCARDPYSSSWNGAFDIW GPGTMVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDT LMISRTPEVTCVVVSVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPSDIAV EWESNGQPENNYLTWPPVLDS DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPG PS3B1921 QSVLTQPPSASGTPGQGVTISCS 1256 NA 1257PSMA_P72_ GSSSNIGSNTVNWFQQLPGTAP E07 KLLIYSDNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCA AWDDSLNGYVFGTGTKVTVLG GSEGKSSGSGSESKSTGGSEVQLVESGGGVVQPGRSLRLSCAAS GFTFITYGMHWVRQAPGKGLE WVAVVSFDESNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAE DTAVYYCARALRDGNNWDYF NGMDVWGQGTTVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFL FPPKPKDTLMISRTPEVTCVVVS VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKA LPAPIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGF YPSDIAVEWESNGQPENNYLT WPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPG PS3B1922 QSVLTQPASVSGSPGQSITISCT 1258NA 1259 PSMA_P72_ GTSSDVGGYNYVSWYQQHPG D01 KAPKLMIYEVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEAD YYCSSYTSSYTYVFGTGTKLTV LGGSEGKSSGSGSESKSTGGSEVQLVESGGDLVQPGGSLRLSCA ASGFTFNNYNMNWVRQAPGK GLEWVSHISTSSSNKYYADSVKGRFSISRDIAKNSMYLQMNSLR DEDTAVYYCARDGVGADYGD YYYYGMD VWGQGTTVTVSSEPKSSDKTHTCPPCPAPEAAGGP SVFLFPPKPKDTLMISRTPEVTC VVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVS NKALPAPIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLV KGFYPSDIAVEWESNGQPENNY LTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPG PS3B1923 QSVLTQPPSVSVAPGQTARITC 1260NA 1261 PSMA_P72_ GGNNSGSKSVHWYQQKPGQAP C01 VLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQ VWDSSSDHGVFGGGTKLTVLG GSEGKSSGSGSESKSTGGSQVQLVESGGGEVQPGRSLRLSCAAS GFSFSGYGMHWVRQAPGKGLE WVAVMSYDGSNRFYVDSVRGRFSISRDNSKNTLYLQMNSLRP EDTAVYYCARDTVWGSHPDAF DIWGQGTVVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKP KDTLMISRTPEVTCVVVSVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPS DIAVEWESNGQPENNYLTWPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG PS3B1924 SYELMQPPSVSVSPGQTARITCS 1262 NA1263 PSMA_P72_ GDALPKQYAYWYQQKPGQAP A10 VLVIYKDSERPSGIPVRFSGSSSGTTVTLTITGVQAEDEADYYCQ SADSSGTYVFGTGTKVTVLGGS EGKSSGSGSESKSTGGSQVQLVESGGGVVQPGRSLRLSCAASGF TFSSYNMNWVRQAPGKGLEW VAIIYYDGSNKYYADSVKGRFTISRDISKNTLYLQMNSLRAEDT AVYYCARERGRDYYGMDVWG QGTTVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDT LMISRTPEVTCVVVSVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPSDIAV EWESNGQPENNYLTWPPVLDS DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPG PS3B1925 QSVLTQPPSASGTPGQRVTISCS 1264 NA 1265PSMA_P70_ GSSSNIGSNTVNWYQQLPGTAP F02 KLLIYSSNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCA AWDDSLNGVVFGGGTKLTVLG GSEGKSSGSGSESKSTGGSEVQLLESGPGLVKPSETLSLTCTVSG GSIISYYWSWIRQPAGKGLEWI GRIYSSGSTNYNPSLKSRVTMSVDTSKNQFSLKLSSVTAADTAV YYCAKVGVWPGAFDIWGQGT MVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMIS RTPEVTCVVVSVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTISKA KGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPSDIAVEWES NGQPENNYLTWPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG PS3B1926 EVQLVESGGGVVQPGRSLRLSC 1266 NA 1267PSMA_P72_ AASGFSFSGYGMHWVRQAPGK G02 GLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSL RVEDTAVYYCARDRIWGSRGY YYGMDVWGQGTTVTVSSGGSEGKSSGSGSESKSTGGSQSALT QPASVSGSPGQSITISCTGASSD VGGYNYVSWYQQHPGKAPKLMIYEVSNRPSGVSNRFSGSKSG NTASLTISGLQAEDEADYYCSS YTITSTLVFGGGTKLTVLEPKSSDKTHTCPPCPAPEAAGGPSVFL FPPKPKDTLMISRTPEVTCVVVS VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKA LPAPIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGF YPSDIAVEWESNGQPENNYLT WPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPG PS3B1927 QVQLVESGGGEVQPGRSLRLSC 1268 NA1269 PSMA_P72_ AASGFSFSGYGMHWVRQAPGK C01 GLEWVAVMSYDGSNRFYVDSVRGRFSISRDNSKNTLYLQMNS LRPEDTAVYYCARDTVWGSHP DAFDIWGQGTVVTVSSGGSEGKSSGSGSESKSTGGSQSVLTQPP SVSVAPGQTARITCGGNNSGSK SVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTI SRVEAGDEADYYCQVWDSSSD HGVFGGGTKLTVLEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKP KDTLMISRTPEVTCVVVSVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPS DIAVEWESNGQPENNYLTWPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG PS3B1928 QVQLQESGGDVVQPGRSLRLS 1270 NA1271 PSMA_P72_ CAASGFSFSGYGLHWVRQAPG lA11 RGLEWVTLISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAKTTVSDPYY YGMDVWGQGTTVTVSSGGSEGKSSGSGSESKSTGGSSYELTQP PSVSVAPGQTARITCGGNNIGS KSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGTNSGNTATL TISRAEAGDEADYYCQVWDSS SDHVVFGGGTKLTVLEPKSSDKTHTCPPCPAPEAAGGPSVFLFPP KPKDTLMISRTPEVTCVVVSVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYP SDIAVEWESNGQPENNYLTWPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG PSMB1041 QSVLTQPPSASGTPGQRVTISCS 1272 NA1273 PSMA_P70_ GSSSNIGSNTVNWYQQLPGTAP F02 KLLIYSSNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCA AWDDSLNGVVFGGGTKLTVLG GSEGKSSGSGSESKSTGGSEVQLLESGPGLVKPSETLSLTCTVSG GSIISYYWSWIRQPAGKGLEWI GRIYSSGSTNYNPSLKSRVTMSVDTSKNQFSLKLSSVTAADTAV YYCAKVGVWPGAFDIWGQGT MVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMIS RTPEVTCVVVSVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTISKA KGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPSDIAVEWES NGQPENNYLTWPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG PSMB1045 SYELMQPPSVSVSPGQTARITCS 1274 NA 1275PSMA_P72_ GDALPKQYAYWYQQKPGQAP A10 VLVIYKDSERPSGIPVRFSGSSSGTTVTLTITGVQAEDEADYYCQ SADSSGTYVFGTGTKVTVLGGS EGKSSGSGSESKSTGGSQVQLVESGGGVVQPGRSLRLSCAASGF TFSSYNMNWVRQAPGKGLEW VAIIYYDGSNKYYADSVKGRFTISRDISKNTLYLQMNSLRAEDT AVYYCARERGRDYYGMDVWG QGTTVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDT LMISRTPEVTCVVVSVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPSDIAV EWESNGQPENNYLTWPPVLDS DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPG PSMB1047 QSVLTQPPSVSVAPGQTARITC 1276 NA 1277PSMA_P72_ GGNNSGSKSVHWYQQKPGQAP C01 VLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQ VWDSSSDHGVFGGGTKLTVLG GSEGKSSGSGSESKSTGGSQVQLVESGGGEVQPGRSLRLSCAAS GFSFSGYGMHWVRQAPGKGLE WVAVMSYDGSNRFYVDSVRGRFSISRDNSKNTLYLQMNSLRP EDTAVYYCARDTVWGSHPDAF DIWGQGTVVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKP KDTLMISRTPEVTCVVVSVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPS DIAVEWESNGQPENNYLTWPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG PSMB1049 QSVLTQPASVSGSPGQSITISCT 1278 NA1279 PSMA_P72_ GTSSDVGGYNYVSWYQQHPG D01 KAPKLMIYEVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEAD YYCSSYTSSYTYVFGTGTKLTV LGGSEGKSSGSGSESKSTGGSEVQLVESGGDLVQPGGSLRLSCA ASGFTFNNYNMNWVRQAPGK GLEWVSHISTSSSNKYYADSVKGRFSISRDIAKNSMYLQMNSLR DEDTAVYYCARDGVGADYGD YYYYGMDVWGQGTTVTVSSEPKSSDKTHTCPPCPAPEAAGGP SVFLFPPKPKDTLMISRTPEVTC VVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVS NKALPAPIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLV KGFYPSDIAVEWESNGQPENNY LTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPG PSMB1051 QSVLTQPPSASGTPGQGVTISCS 1280NA 1281 PSMA_P72_ GSSSNIGSNTVNWFQQLPGTAP E07 KLLIYSDNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCA AWDDSLNGYVFGTGTKVTVLG GSEGKSSGSGSESKSTGGSEVQLVESGGGVVQPGRSLRLSCAAS GFTFITYGMHWVRQAPGKGLE WVAVVSFDESNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAE DTAVYYCARALRDGNNWDYF NGMDVWGQGTTVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFL FPPKPKDTLMISRTPEVTCVVVS VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKA LPAPIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGF YPSDIAVEWESNGQPENNYLT WPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPG PSMB1052 SYELTQPPSVSVAPGQTARITCG 1282NA 1283 PSMA_P72_ GNNIGSKSVHWYQQKPGQAPV F07 LVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQV WDSSTDHVVFGGGTKLTVLGG SEGKSSGSGSESKSTGGSEVQLVESGGGVVQPGRSLRLSCAASG FTFSSYGMNWVRQAPGKGLEW VAVTSYDGSNKYYADSVKGRFTISRDISKNTLYLQMSSLRAEDT AVYYCARDPYSSSWNGAFDIW GPGTMVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDT LMISRTPEVTCVVVSVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPSDIAV EWESNGQPENNYLTWPPVLDS DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPG PSMB1060 EIVLTQSPGTLSVSPGERATLSC 1284 NA 1285PSMA_P75_ RASQSVRSNLAWYQQKPGQAP F01 RLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCHQ YNDWPPYTFGQGTKLEIKGGSE GKSSGSGSESKSTGGSQVQLQESGGGVVQPGRSLRLSCAASGFT FSTYGMHWVRQAPGKGLEWV AFISYDGSNKYYADSVKGRFTISRDNSKHTLYLQMNSLRAEDT AVYYCAGRDNLRFLEWFMDV WGQGTTVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKD TLMISRTPEVTCVVVSVSHEDP EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPSDI AVEWESNGQPENNYLTWPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPG PSMB1068 QVQLQESGGDVVQPGRSLRLS 1286 NA 1287PSMA_P72_ CAASGFSFSGYGLHWVRQAPG A11 RGLEWVTLISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAKTTVSDPYY YGMDVWGQGTTVTVSSGGSEGKSSGSGSESKSTGGSSYELTQP PSVSVAPGQTARITCGGNNIGS KSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGTNSGNTATL TISRAEAGDEADYYCQVWDSS SDHVVFGGGTKLTVLEPKSSDKTHTCPPCPAPEAAGGPSVFLFPP KPKDTLMISRTPEVTCVVVSVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYP SDIAVEWESNGQPENNYLTWPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG PSMB1069 QVQLVESGGGEVQPGRSLRLSC 1288 NA1289 PSMA_P72_ AASGFSFSGYGMHWVRQAPGK C01 GLEWVAVMSYDGSNRFYVDSVRGRFSISRDNSKNTLYLQMNS LRPEDTAVYYCARDTVWGSHP DAFDIWGQGTVVTVSSGGSEGKSSGSGSESKSTGGSQSVLTQPP SVSVAPGQTARITCGGNNSGSK SVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTI SRVEAGDEADYYCQVWDSSSD HGVFGGGTKLTVLEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKP KDTLMISRTPEVTCVVVSVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGFYPS DIAVEWESNGQPENNYLTWPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG PSMB1075 EVQLVESGGGVVQPGRSLRLSC 1290 NA1291 PSMA_P72_ AASGFSFSGYGMHWVRQAPGK G02 GLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSL RVEDTAVYYCARDRIWGSRGY YYGMDVWGQGTTVTVSSGGSEGKSSGSGSESKSTGGSQSALT QPASVSGSPGQSITISCTGASSD VGGYNYVSWYQQHPGKAPKLMIYEVSNRPSGVSNRFSGSKSG NTASLTISGLQAEDEADYYCSS YTITSTLVFGGGTKLTVLEPKSSDKTHTCPPCPAPEAAGGPSVFL FPPKPKDTLMISRTPEVTCVVVS VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKA LPAPIEKTISKAKGQPREPQVYVLPPSREEMTKNQVSLLCLVKGF YPSDIAVEWESNGQPENNYLT WPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPG PSMB290 QSVLTQPASVSGSPGQSITISCT 1292 NA1293 PSMA_P72_ 8 GTSSDVGGYNYVSWYQQHPG D01-HC- KAPKLMIYEVSNRPSGVSNRFSD95E GSKSGNTASLTISGLQAEDEAD YYCSSYTSSYTYVFGTGTKLTVLGGSEGKSSGSGSESKSTGGSE VQLVESGGDLVQPGGSLRLSCA ASGFTFNNYNMNWVRQAPGKGLEWVSHISTSSSNKYYADSVK GRFSISRDIAKNSMYLQMNSLR DEDTAVYYCAREGVGADYGDYYYYGMDVWGQGTTVTVSSE PKSSDKTHTCPPCPAPEAAGGP SVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYVLPPSREEMTKNQVSLLCLV KGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPG PSMB290SYELMQPPSVSVSPGQTARITCS 1294 NA 1295 PSMA_P72_ 9 GDALPKQYAYWYQQKPGQAPA10-HC- VLVIYKDSERPSGIPVRFSGSSS G54E GTTVTLTITGVQAEDEADYYCQSADSSGTYVFGTGTKVTVLGGS EGKSSGSGSESKSTGGSQVQLV ESGGGVVQPGRSLRLSCAASGFTFSSYNMNWVRQAPGKGLEW VAIIYYDESNKYYADSVKGRFT ISRDISKNTLYLQMNSLRAEDTAVYYCARERGRDYYGMDVWG QGTTVTVSSEPKSSDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPE VKFNWYVDGVEVHNAKTKPR EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYVLPPSREE MTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDS DGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLS LSPGPS3B1391 EVQLVESGGGLVKPGGSLRLSC 1459 EIVMTQSPGTLS 1460 PSMHB49SVASGFTFSFYSMNWVRQAPGK LSPGERATLSCR C1133_011 GLDWVSSISSSGNYIYYADSVKASQSVSSSFLAW A11_1 GRFTISRDNAKNSLHLHMNSLK YQQKPGQAPRLAEDTAMYFCARSYSGSYDAFD LISGASSRATGIP FWGQGTMVTVSSASTKGPSVF DRFSVSGSGTDFPLAPSSKSTSGGTAALGCLVKD TLTISRLEPEDFA YFPEPVTVSWNSGALTSGVHTF VYYCQQYGVSPPAVLQSSGLYSLSSVVTVPSSSL WTFGQGTKVEIK GTQTYICNVNHKPSNTKVDKK RTVAAPSVFIFPPVEPKSCDKTHTCPPCPAPEAAG SDEQLKSGTASV GPSVFLFPPKPKDTLMISRTPEV VCLLNNFYPREATCVVVSVSHEDPEVKFNWYVD KVQWKVDNAL GVEVHNAKTKPREEQYNSTYR QSGNSQESVTEQVVSVLTVLHQDWLNGKEYKC DSKDSTYSLSST KVSNKALPAPIEKTISKAKGQP LTLSKADYEKHREPQVYTLPPSREEMTKNQVSL KVYACEVTHQG SCAVKGFYPSDIAVEWESNGQP LSSPVTKSFNRGENNYKTTPPVLDSDGSFFLVSK EC LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGKPS3B1396 EVQLVESGGGLVQPGGSLRLSC 1461 QSVLTQPPSVSA 1462 PSMB896-AASGFTFSSYAMSWVRQAPGK APGQKVTISCSG G100A GLEWVSAISGGIGSTYYADSVKSSSNIGINYVSW GRFTISRDNSKNTLWLQMNSLR YQQLPGTAPKLL AEDTAVYYCAKDAVGATPYYFIYDNNKRPSGIP DYWGQGTLVTVSSASTKGPSV DRFSGSKSGTSA FPLAPSSKSTSGGTAALGCLVKTLGITGLQTGDE DYFPEPVTVSWNSGALTSGVHT ADYYCGTWDSS FPAVLQSSGLYSLSSVVTVPSSSLSAVVFGGGTKL LGTQTYICNVNHKPSNTKVDK TVLGQPKAAPSV KVEPKSCDKTHTCPPCPAPEAATLFPPSSEELQAN GGPSVFLFPPKPKDTLMISRTPE KATLVCLISDFY VTCVVVSVSHEDPEVKFNWYVPGAVTVAWKAD DGVEVHNAKTKPREEQYNSTY SSPVKAGVETTT RVVSVLTVLHQDWLNGKEYKCPSKQSNNKYAAS KVSNKALPAPIEKTISKAKGQP SYLSLTPEQWKS REPQVYTLPPSREEMTKNQVSLHRSYSCQVTHEG SCAVKGFYPSDIAVEWESNGQP STVEKTVAPTEC ENNYKTTPPVLDSDGSFFLVSKS LTVDKSRWQQGNVFSCSVMHE ALHNRFTQKSLSLSPGK

TABLE 61 PSMA × CD3 Bispecific Antibodies: Clone Descriptions NameBispecific Description PS3B917 FAB-A: CD3B891 (CD3B376 − K477); FAB-B:PSMB889 PS3B918 FAB-A: CD3B891 (CD3B376 − K477); FAB-B: PSMB890 PS3B913FAB-A: CD3B891 (CD3B376 − K477); FAB-B: PSMB891 PS3B915 FAB-A: CD3B891(CD3B376 − K477); FAB-B: PSMB892 PS3B914 FAB-A: CD3B891 (CD3B376 −K477); FAB-B: PSMB893 PS3B916 FAB-A: CD3B891 (CD3B376 − K477); FAB-B:PSMB894 PS3B919 FAB-A: CD3B891 (CD3B376 − K477); FAB-B: PSMB895 PS3B921FAB-A: CD3B891 (CD3B376 − K477); FAB-B: PSMB896 PS3B920 FAB-A: CD3B891(CD3B376 − K477); FAB-B: PSMB897 PS3B922 FAB-A: CD3B891 (CD3B376 −K477); FAB-B: PSMB898 PS3B912 FAB-A: CD3B891 (CD3B376 − K477); FAB-B:PSMB899 PS3B930 FAB-A: CD3B2183 (CD3W245); FAB-B: PSMB889 PS3B931 FAB-A:CD3B2183 (CD3W245); FAB-B: PSMB890 PS3B926 FAB-A: CD3B2183 (CD3W245);FAB-B: PSMB891 PS3B928 FAB-A: CD3B2183 (CD3W245); FAB-B: PSMB892 PS3B927FAB-A: CD3B2183 (CD3W245); FAB-B: PSMB893 PS3B929 FAB-A: CD3B2183(CD3W245); FAB-B: PSMB894 PS3B932 FAB-A: CD3B2183 (CD3W245); FAB-B:PSMB895 PS3B934 FAB-A: CD3B2183 (CD3W245); FAB-B: PSMB896 PS3B933 FAB-A:CD3B2183 (CD3W245); FAB-B: PSMB897 PS3B935 FAB-A: CD3B2183 (CD3W245);FAB-B: PSMB898 PS3B925 FAB-A: CD3B2183 (CD3W245); FAB-B: PSMB899PS3B1352 FAB-A: CD3B2197 (CD3B376 + K477); FAB-B: PSMB946 CD3B2197 isCD3B376 but with C-term K PSMB946 is PSMB895 but with C-term K PS3B1353FAB-A: CD3B2197 (CD3B376 + K477); FAB-B: PSMB947 CD3B2197 is CD3B376 butwith C-term K PSMB947 is PSMB896 but with C-term K PS3B1354 FAB-A:CD3B2197 (CD3B376 + K477); FAB-B: PSMB948 CD3B2197 is CD3B376 but withC-term K PSMB948 is PSMB897 but with C-term K PS3B1355 FAB-A: CD3B2197(CD3B376 + K477); FAB-B: PSMB949 CD3B2197 is CD3B376 with C-term KPSMB949 is PSMB898 but with C-term K PS3B1356 FAB-A: CD3B2200 (CD3B450 +K477); FAB-B: PSMB946 CD3B2200 is CD3B450 with the C-term K PSMB946 isPSMB895 but with C-term K PS3B1357 FAB-A: CD3B2200 (CD3B450 + K477);FAB-B: PSMB947 CD3B2200 is CD3B450 with the C-term K PSMB947 is PSMB896but with C-term K PS3B1358 FAB-A: CD3B2200 (CD3B450 + K477); FAB-B:PSMB949 CD3B2200 is CD3B450 with the C-term K PSMB949 is PSMB898 butwith C-term K PSMB937 FAB-A: CD3B2186 (CD3B450 − K477); FAB-B: PSMB897(PSMB948) CD3B2186 is CD3B450 without the C-term K PSMB948 is PSMB897but with C-term K

TABLE 62 PSMA × CD3 Bispecific Antibodies: CD3 Arm Descriptions SEQ SEQID ID CD3 Arm Name HC1 NO. LC1 NO. Description PS3QVQLQQSGPRLVRPSQTLSLTCAI 1296 QSALTQPASVS 1297 with CD3B376 B917SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891 EWLGRTYYRSKWLYDYAVSVKSGTSSNIGTYKFV without K477 or RITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKACD3B2197 with TALYYCARGYSSSFDYWGQGTL PKVLLYEVSKR K477 in HC1)VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSK HC1 ConstructGGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID: NSGALTSGVHTFPAVLQSSGLYSQAEDQADYHC PBD000100300 LSSVVTVPSSSLGTQTYICNVNHK VSYAGSGTLLFLC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1298 QSALTQPASVS 1299with CD3B376 B918 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000100300 LSSVVTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1300 QSALTQPASVS 1301with CD3B376 B913 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000100300 LSSVVTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1302 QSALTQPASVS 1303with CD3B376 B915 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000100300 LSSVVTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1304 QSALTQPASVS 1305with CD3B376 B914 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000100300 LSSVVTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1306 QSALTQPASVS 1307with CD3B376 B916 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000100300 LSSVVTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1308 QSALTQPASVS 1309with CD3B376 B919 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000100300 LSSWTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1310 QSALTQPASVS 1311with CD3B376 B921 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000100300 LSSWTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1312 QSALTQPASVS 1313with CD3B376 B920 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000100300 LSSVVTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1314 QSALTQPASVS 1315with CD3B376 B922 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000100300 LSSWTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1316 QSALTQPASVS 1317with CD3B376 B912 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000100300 LSSWTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000044707MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQWTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPG ECS PS3 EVQLVESGGGLVKPGGSLRLSCA 1318 DIQMTQSPSSLS 1319with CD3W245 B930 ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3EVQLVESGGGLVKPGGSLRLSCA 1320 DIQMTQSPSSLS 1321 with CD3W245 B931ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3EVQLVESGGGLVKPGGSLRLSCA 1322 DIQMTQSPSSLS 1323 with CD3W245 B926ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3EVQLVESGGGLVKPGGSLRLSCA 1324 DIQMTQSPSSLS 1325 with CD3W245 B928ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3EVQLVESGGGLVKPGGSLRLSCA 1326 DIQMTQSPSSLS 1327 with CD3W245 B927ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3EVQLVESGGGLVKPGGSLRLSCA 1328 DIQMTQSPSSLS 1329 with CD3W245 B929ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3EVQLVESGGGLVKPGGSLRLSCA 1330 DIQMTQSPSSLS 1331 with CD3W245 B932ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3EVQLVESGGGLVKPGGSLRLSCA 1332 DIQMTQSPSSLS 1333 with CD3W245 B934ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3EVQLVESGGGLVKPGGSLRLSCA 1334 DIQMTQSPSSLS 1335 with CD3W245 B933ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3EVQLVESGGGLVKPGGSLRLSCA 1336 DIQMTQSPSSLS 1337 with CD3W245 B935ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3EVQLVESGGGLVKPGGSLRLSCA 1338 DIQMTQSPSSLS 1339 with CD3W245 B925ASGFTFSRYNMNWVRQAPGKGL ASVGDRVTITC arm (CD3B2183EWVSSISTSSNYIYYADSVKGRFT RARQSIGTAIH without K477)FSRDNAKNSLDLQMSGLRAEDT WYQQKPGKAP HC1 Construct AIYYCTRGWGPFDYWGQGTLVTKLLIKYASESIS ID: VSSASTKGPSVFPLAPSSKSTSGG GVPSRFSGSGS PBD000100302TAALGCLVKDYFPEPVTVSWNS GTDFTLTISSLQ LC1 ContructGALTSGVHTFPAVLQSSGLYSLSS PEDFATYYCQQ ID: VVTVPSSSLGTQTYICNVNHKPSSGSWPYTFGQG PBD000084982 NTKVDKKVEPKSCDKTHTCPPCP TKLEIKRTVAAAPEAAGGPSVFLFPPKPKDTLMIS PSVFIFPPSDEQ RTPEVTCVVVSVSHEDPEVKFNWLKSGTASVVCL YVDGVEVHNAKTKPREEQYNST LNNFYPREAKV YRVVSVLTVLHQDWLNGKEYKCQWKVDNALQS KVSNKALPAPIEKTISKAKGQPRE GNSQESVTEQD PQVYTLPPSREEMTKNQVSLTCLSKDSTYSLSSTL VKGFYPSDIAVEWESNGQPENNY TLSKADYEKHK KTTPPVLDSDGSFLLYSKLTVDKVYACEVTHQGL SRWQQGNVFSCSVMHEALHNHY SSPVTKSFNRGE TQKSLSLSP C PS3QVQLQQSGPRLVRPSQTLSLTCAI 1340 QSALTQPASVS 1341 with CD3B376 B135SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891 2EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000108469 LSSWTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000108469MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPGK ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1342 QSALTQPASVS 1343with CD3B376 B135 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891 3EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000108469 LSSWTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000108469MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPGK ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1344 QSALTQPASVS 1345with CD3B376 B135 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891 4EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000108469 LSSVVTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000108469MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPGK ECS PS3 QVQLQQSGPRLVRPSQTLSLTCAI 1346 QSALTQPASVS 1347with CD3B376 B135 SGDSVFNNNAAWSWIRQSPSRGL GSPGQSITISCT arm (CD3B891 5EWLGRTYYRSKWLYDYAVSVKS GTSSNIGTYKFV without K477 orRITVNPDTSRNQFTLQLNSVTPED SWYQQHPDKA CD3B2197 with TALYYCARGYSSSFDYWGQGTLPKVLLYEVSKR K477 in HC1) VTVSSASTKGPSVFPLAPSSKSTS PSGVSSRFSGSKHC1 Construct GGTAALGCLVKDYFPEPVTVSW SGNTASLTISGL ID:NSGALTSGVHTFPAVLQSSGLYS QAEDQADYHC PBD000108469 LSSVVTVPSSSLGTQTYICNVNHKVSYAGSGTLLF LC1 Contruct PSNTKVDKKVEPKSCDKTHTCPP GGGTKLTVLGQ ID:CPAPEAAGGPSVFLFPPKPKDTL PKAAPSVTLFPP PBD000108469MISRTPEVTCVVVSVSHEDPEVK SSEELQANKAT FNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGANSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSP YKCKVSNKALPAPIEKTISKAKG VKAGVETTTPSQPREPQVYTLPPSREEMTKNQVS KQSNNKYAASS LTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKSENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHE TVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPTHNHYTQKSLSLSPGK ECS PS3 QVQLQQSGPGLVKPSQTLSLTCA 1348 QSALTQPASVS 1349with CD3B450 B135 ISGDSVFNNNAAWSWIRQSPSRG GSPGQSITISCT arm (CD3B2186 6LEWLGRTYYRSKWLYDYAVSVK GTSSNIGTYKFV without K477 orSRITINPDTSKNQFSLQLNSVTPE SWYQQHPGKA CD3B2200 with DTAVYYCARGYSSSFDYWGQGTPKVMIYEVSKR K477 in HC1) LVTVSSASTKGPSVFPLAPSSKST PSGVSNRFSGSHC1 Construct SGGTAALGCLVKDYFPEPVTVS KSGNTASLTISG ID:WNSGALTSGVHTFPAVLQSSGLY LQAEDEADYYC PBD000108470SLSSVVTVPSSSLGTQTYICNVNH VSYAGSGTLLF LC1 ContructKPSNTKVDKKVEPKSCDKTHTCP GGGTKLTVLGQ ID: PCPAPEAAGGPSVFLFPPKPKDTLPKAAPSVTLFPP PBD000108470 MISRTPEVTCVVVSVSHEDPEVK SSEELQANKATFNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGA NSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSPYKCKVSNKALPAPIEKTISKAKG VKAGVETTTPS QPREPQVYTLPPSREEMTKNQVS KQSNNKYAASSLTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKS ENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHETVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPT HNHYTQKSLSLSPGK ECS PS3QVQLQQSGPGLVKPSQTLSLTCA 1350 QSALTQPASVS 1351 with CD3B450 B135ISGDSVFNNNAAWSWIRQSPSRG GSPGQSITISCT arm (CD3B2186 7LEWLGRTYYRSKWLYDYAVSVK GTSSNIGTYKFV without K477 orSRITINPDTSKNQFSLQLNSVTPE SWYQQHPGKA CD3B2200 with DTAVYYCARGYSSSFDYWGQGTPKVMIYEVSKR K477 in HC1) LVTVSSASTKGPSVFPLAPSSKST PSGVSNRFSGSHC1 Construct SGGTAALGCLVKDYFPEPVTVS KSGNTASLTISG ID:WNSGALTSGVHTFPAVLQSSGLY LQAEDEADYYC PBD000108470SLSSVVTVPSSSLGTQTYICNVNH VSYAGSGTLLF LC1 ContructKPSNTKVDKKVEPKSCDKTHTCP GGGTKLTVLGQ ID: PCPAPEAAGGPSVFLFPPKPKDTLPKAAPSVTLFPP PBD000108470 MISRTPEVTCVVVSVSHEDPEVK SSEELQANKATFNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGA NSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSPYKCKVSNKALPAPIEKTISKAKG VKAGVETTTPS QPREPQVYTLPPSREEMTKNQVS KQSNNKYAASSLTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKS ENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHETVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPT HNHYTQKSLSLSPGK ECS PS3QVQLQQSGPGLVKPSQTLSLTCA 1352 QSALTQPASVS 1353 with CD3B450 B135ISGDSVFNNNAAWSWIRQSPSRG GSPGQSITISCT arm (CD3B2186 8LEWLGRTYYRSKWLYDYAVSVK GTSSNIGTYKFV without K477 orSRITINPDTSKNQFSLQLNSVTPE SWYQQHPGKA CD3B2200 with DTAVYYCARGYSSSFDYWGQGTPKVMIYEVSKR K477 in HC1) LVTVSSASTKGPSVFPLAPSSKST PSGVSNRFSGSHC1 Construct SGGTAALGCLVKDYFPEPVTVS KSGNTASLTISG ID:WNSGALTSGVHTFPAVLQSSGLY LQAEDEADYYC PBD000108470SLSSVVTVPSSSLGTQTYICNVNH VSYAGSGTLLF LC1 ContructKPSNTKVDKKVEPKSCDKTHTCP GGGTKLTVLGQ ID: PCPAPEAAGGPSVFLFPPKPKDTLPKAAPSVTLFPP PBD000108470 MISRTPEVTCVVVSVSHEDPEVK SSEELQANKATFNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGA NSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSPYKCKVSNKALPAPIEKTISKAKG VKAGVETTTPS QPREPQVYTLPPSREEMTKNQVS KQSNNKYAASSLTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKS ENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHETVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPT HNHYTQKSLSLSPGK ECS PSMQVQLQQSGPGLVKPSQTLSLTCA 1354 QSALTQPASVS 1355 with CD3B450 B937ISGDSVFNNNAAWSWIRQSPSRG GSPGQSITISCT arm (CD3B2186LEWLGRTYYRSKWLYDYAVSVK GTSSNIGTYKFV without K477 orSRITINPDTSKNQFSLQLNSVTPE SWYQQHPGKA CD3B2200 with DTAVYYCARGYSSSFDYWGQGTPKVMIYEVSKR K477 in HC1) LVTVSSASTKGPSVFPLAPSSKST PSGVSNRFSGSHC1 Construct SGGTAALGCLVKDYFPEPVTVS KSGNTASLTISG ID:WNSGALTSGVHTFPAVLQSSGLY LQAEDEADYYC PBD000100305SLSSVVTVPSSSLGTQTYICNVNH VSYAGSGTLLF LC1 ContructKPSNTKVDKKVEPKSCDKTHTCP GGGTKLTVLGQ ID: PCPAPEAAGGPSVFLFPPKPKDTLPKAAPSVTLFPP PBD000045576 MISRTPEVTCVVVSVSHEDPEVK SSEELQANKATFNWYVDGVEVHNAKTKPREEQY LVCLISDFYPGA NSTYRVVSVLTVLHQDWLNGKE VTVAWKADSSPYKCKVSNKALPAPIEKTISKAKG VKAGVETTTPS QPREPQVYTLPPSREEMTKNQVS KQSNNKYAASSLTCLVKGFYPSDIAVEWESNGQP YLSLTPEQWKS ENNYKTTPPVLDSDGSFLLYSKL HRSYSCQVTHETVDKSRWQQGNVFSCSVMHEAL GSTVEKTVAPT HNHYTQKSLSLSPG ECS

TABLE 63 PSMA × CD3 Bispecific Antibodies: PSMA Arm Descriptions SEQ SEQPSMA ID ID Arm Name HC2 NO. LC2 NO. Description PS3BEVQLVESGGGLVKPGGSLRLSCAAS 1356 QSVLTQPPSVSG 1357 PSMB889 917GFTFSRYNMNWVRQAPGKGLEWV APGQRVTISCTG HC2 SSINSNSRYIYYADSVKGRFTISRDSSSFNLGAGYDV Construct AKNSLYLQMNSLRAEDTAVYYCAK HWYQQVPGTVP ID:TMGDYYYYYGMDVWGQGTTVTVS KLLIYDNSNRPS PBD000101SASTKGPSVFPLAPSSKSTSGGTAAL GVPDRFSGSKSG 312 GCLVKDYFPEPVTVSWNSGALTSGTSASLAITGLQA VHTFPAVLQSSGLYSLSSVVTVPSSS EDETVYYCQSYLGTQTYICNVNHKPSNTKVDKKVEP DSSLSGVVFGGG KSCDKTHTCPPCPAPEAAGGPSVFLTKLTVLGQPKA FPPKPKDTLMISRTPEVTCVVVSVSH APSVTLFPPSSEEEDPEVKFNWYVDGVEVHNAKTKPR LQANKATLVCLI EEQYNSTYRVVSVLTVLHQDWLNGSDFYPGAVTVA KEYKCKVSNKALPAPIEKTISKAKG WKADSSPVKAGQPREPQVYTLPPSREEMTKNQVSLT VETTTPSKQSNN CLVKGFYPSDIAVEWESNGQPENNYKYAASSYLSLTP KTTPPVLDSDGSFFLYSRLTVDKSR EQWKSHRSYSCWQQGNVFSCSVMHEALHNHYTQK QVTHEGSTVEKT SLSLSPG VAPTECS PS3BEVQLVESGGGLVKPGGSLRLSCAAS 1358 SSELTQPPSVSG 1359 PSMB890 918GFTFSRYNMNWVRQAPGKGLEWV APGQRVTISCAG HC2 SSINSNSRYIYYADSVKGRFTISRDSSLSNIGAGYDVH Construct AKNSLYLQMNSLRAEDTAVYYCAK WYQQLPGTAPK ID:TMGDYYYYYGMDVWGQGTTVTVS LLIYGNINRLSG PBD000101SASTKGPSVFPLAPSSKSTSGGTAAL VPERFSGSKSGT 312 GCLVKDYFPEPVTVSWNSGALTSGSASLAITGLQAE VHTFPAVLQSSGLYSLSSVVTVPSSS DGADYYCQSYDLGTQTYICNVNHKPSNTKVDKKVEP SSLSSYVFGTGT KSCDKTHTCPPCPAPEAAGGPSVFLKVTVLGQPKAA FPPKPKDTLMISRTPEVTCVVVSVSH PSVTLFPPSSEELEDPEVKFNWYVDGVEVHNAKTKPR QANKATLVCLIS EEQYNSTYRVVSVLTVLHQDWLNGDFYPGAVTVAW KEYKCKVSNKALPAPIEKTISKAKG KADSSPVKAGVQPREPQVYTLPPSREEMTKNQVSLT etttpskqsnnk CLVKGFYPSDIAVEWESNGQPENNYYAASSYLSLTPE KTTPPVLDSDGSFFLYSRLTVDKSR QWKSHRSYSCQWQQGNVFSCSVMHEALHNHYTQK VTHEGSTVEKTV SLSLSPG APTECS PS3BEVQLVESGGGVVQPGRSLRLSCAAS 1360 QSVLTQPPSASG 1361 PSMB891 913GFTFITYGMHWVRQAPGKGLEWVA TPGQGVTISCSG HC2 VVSFDESNKYYADSVKGRFTISRDNSSSNIGSNTVNW Construct SKNTLYLQMNSLRAEDTAVYYCAR FQQLPGTAPKLL ID:ALRDGNNWDYFNGMDVWGQGTT IYSDNQRPSGVP PBD000101 VTVSSASTKGPSVFPLAPSSKSTSGGDRFSGSKSGTSA 316 TAALGCLVKDYFPEPVTVSWNSGA SLAISGLQSEDELTSGVHTFPAVLQSSGLYSLSSVVT ADYYCAAWDDS VPSSSLGTQTYICNVNHKPSNTKVDLNGYVFGTGTK KKVEPKSCDKTHTCPPCPAPEAAGG VTVLGQPKAAPSPSVFLFPPKPKDTLMISRTPEVTCVV VTLFPPSSEELQA VSVSHEDPEVKFNWYVDGVEVHNANKATLVCLISDF KTKPREEQYNSTYRVVSVLTVLHQ YPGAVTVAWKADWLNGKEYKCKVSNKALPAPIEKTI DSSPVKAGVETT SKAKGQPREPQVYTLPPSREEMTKNTPSKQSNNKYA QVSLTCLVKGFYPSDIAVEWESNGQ ASSYLSLTPEQWPENNYKTTPPVLDSDGSFFLYSRLT KSHRSYSCQVTH VDKSRWQQGNVFSCSVMHEALHNEGSTVEKTVAPT HYTQKSLSLSPG ECS PS3B EVQLVESGGGVVQPGRSLRLSCAAS 1362QSVLTQPPSVSG 1363 PSMB892 915 GFTFITYGMHWVRQAPGKGLEWVA APGQRVTISCTG HC2VVSFDESNKYYADSVKGRFTISRDN SSSNIGADYDVH ConstructSKNTLYLQMNSLRAEDTAVYYCAR WYQHLPGTAPK ID: ALRDGNNWDYFNGMDVWGQGTTLLIYGNSNRPSG PBD000101 VTVSSASTKGPSVFPLAPSSKSTSGG VPDRFSGSKSGT 316TAALGCLVKDYFPEPVTVSWNSGA SASLAITGLQAE LTSGVHTFPAVLQSSGLYSLSSVVTDETDYYCQSYD VPSSSLGTQTYICNVNHKPSNTKVD SSLSGWVFGGGTKKVEPKSCDKTHTCPPCPAPEAAGG KLTVLGQPKAAP PSVFLFPPKPKDTLMISRTPEVTCVVSVTLFPPSSEELQ VSVSHEDPEVKFNWYVDGVEVHNA ANKATLVCLISDKTKPREEQYNSTYRVVSVLTVLHQ FYPGAVTVAWK DWLNGKEYKCKVSNKALPAPIEKTIADSSPVKAGVET SKAKGQPREPQVYTLPPSREEMTKN TTPSKQSNNKYAQVSLTCLVKGFYPSDIAVEWESNGQ ASSYLSLTPEQW PENNYKTTPPVLDSDGSFFLYSRLTKSHRSYSCQVTH VDKSRWQQGNVFSCSVMHEALHN EGSTVEKTVAPT HYTQKSLSLSPG ECS PS3BQVQLVESGGGVVQPGRSLRLSCVA 1364 QSVLTQPPSASG 1365 PSMB893 914SGFTFSSYGIHWVRQAPGKGLEWV TPGQGVTISCSG HC2 AVIWYDGSNKYYADSVKGRFTISRSSSNIGSNTVNW Construct DNSKNTLYLQMNSLRAEDTAVYYS FQQLPGTAPKLL ID:VRGVGPTSYYYNYGMDVWGQGTT IYSDNQRPSGVP PBD000101VTVSSASTKGPSVFPLAPSSKSTSGG DRFSGSKSGTSA 318 TAALGCLVKDYFPEPVTVSWNSGASLAISGLQSEDE LTSGVHTFPAVLQSSGLYSLSSVVT ADYYCAAWDDSVPSSSLGTQTYICNVNHKPSNTKVD LNGYVFGTGTK KKVEPKSCDKTHTCPPCPAPEAAGGVTVLGQPKAAPS PSVFLFPPKPKDTLMISRTPEVTCVV VTLFPPSSEELQAVSVSHEDPEVKFNWYVDGVEVHNA NKATLVCLISDF KTKPREEQYNSTYRVVSVLTVLHQYPGAVTVAWKA DWLNGKEYKCKVSNKALPAPIEKTI DSSPVKAGVETTSKAKGQPREPQVYTLPPSREEMTKN TPSKQSNNKYA QVSLTCLVKGFYPSDIAVEWESNGQASSYLSLTPEQW PENNYKTTPPVLDSDGSFFLYSRLT KSHRSYSCQVTHVDKSRWQQGNVFSCSVMHEALHN EGSTVEKTVAPT HYTQKSLSLSPG ECS PS3BQVQLVESGGGVVQPGRSLRLSCVA 1366 QSVLTQPPSVSG 1367 PSMB894 916SGFTFSSYGIHWVRQAPGKGLEWV APGQRVTISCTG HC2 AVIWYDGSNKYYADSVKGRFTISRSSSNIGADYDVH Construct DNSKNTLYLQMNSLRAEDTAVYYS WYQHLPGTAPK ID:VRGVGPTSYYYNYGMDVWGQGTT LLIYGNSNRPSG PBD000101VTVSSASTKGPSVFPLAPSSKSTSGG VPDRFSGSKSGT 318 TAALGCLVKDYFPEPVTVSWNSGASASLAITGLQAE LTSGVHTFPAVLQSSGLYSLSSVVT DETDYYCQSYDVPSSSLGTQTYICNVNHKPSNTKVD SSLSGWVFGGGT KKVEPKSCDKTHTCPPCPAPEAAGGKLTVLGQPKAAP PSVFLFPPKPKDTLMISRTPEVTCVV SVTLFPPSSEELQVSVSHEDPEVKFNWYVDGVEVHNA ANKATLVCLISD KTKPREEQYNSTYRVVSVLTVLHQFYPGAVTVAWK DWLNGKEYKCKVSNKALPAPIEKTI ADSSPVKAGVETSKAKGQPREPQVYTLPPSREEMTKN TTPSKQSNNKYA QVSLTCLVKGFYPSDIAVEWESNGQASSYLSLTPEQW PENNYKTTPPVLDSDGSFFLYSRLT KSHRSYSCQVTHVDKSRWQQGNVFSCSVMHEALHN EGSTVEKTVAPT HYTQKSLSLSPG ECS PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1368 QSVLTQPPSVSA 1369 PSMB895 919GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGIGSTYYADSVKGRFTISRDNSSSSNIGNNYVSW Construct KNTLWLQMNSLRAEDTAVYYCAK YQQLPGTAPKLL ID:DGVGATPYYFDYWGQGTLVTVSSA IYDNNKRPSGIP PBD000101STKGPSVFPLAPSSKSTSGGTAALGC DRFSGSKSGTSA 320 LVKDYFPEPVTVSWNSGALTSGVHTLGITGLQTGDE TFPAVLQSSGLYSLSSVVTVPSSSLG ADYYCGTWDSSTQTYICNVNHKPSNTKVDKKVEPKS LSAYVFGTGTKV CDKTHTCPPCPAPEAAGGPSVFLFPPTVLGQPKAAPSV KPKDTLMISRTPEVTCVVVSVSHED TLFPPSSEELQANPEVKFNWYVDGVEVHNAKTKPREE KATLVCLISDFY QYNSTYRVVSVLTVLHQDWLNGKEPGAVTVAWKAD YKCKVSNKALPAPIEKTISKAKGQP SSPVKAGVETTTREPQVYTLPPSREEMTKNQVSLTCL PSKQSNNKYAAS VKGFYPSDIAVEWESNGQPENNYKSYLSLTPEQWKS TTPPVLDSDGSFFLYSRLTVDKSRW HRSYSCQVTHEGQQGNVFSCSVMHEALHNHYTQKSL STVEKTVAPTEC SLSPG S PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1370 QSVLTQPPSVSA 1371 PSMB896 921GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGIGSTYYADSVKGRFTISRDNSSSSNIGINYVSW Construct KNTLWLQMNSLRAEDTAVYYCAK YQQLPGTAPKLL ID:DGVGATPYYFDYWGQGTLVTVSSA IYDNNKRPSGIP PBD000101STKGPSVFPLAPSSKSTSGGTAALGC DRFSGSKSGTSA 320 LVKDYFPEPVTVSWNSGALTSGVHTLGITGLQTGDE TFPAVLQSSGLYSLSSVVTVPSSSLG ADYYCGTWDSSTQTYICNVNHKPSNTKVDKKVEPKS LSAVVFGGGTKL CDKTHTCPPCPAPEAAGGPSVFLFPPTVLGQPKAAPSV KPKDTLMISRTPEVTCVVVSVSHED TLFPPSSEELQANPEVKFNWYVDGVEVHNAKTKPREE KATLVCLISDFY QYNSTYRVVSVLTVLHQDWLNGKEPGAVTVAWKAD YKCKVSNKALPAPIEKTISKAKGQP SSPVKAGVETTTREPQVYTLPPSREEMTKNQVSLTCL PSKQSNNKYAAS VKGFYPSDIAVEWESNGQPENNYKSYLSLTPEQWKS TTPPVLDSDGSFFLYSRLTVDKSRW HRSYSCQVTHEGQQGNVFSCSVMHEALHNHYTQKSL STVEKTVAPTEC SLSPG s PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1372 QSVLTQPPSVSA 1373 PSMB897 920GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGSGSTYYADSVKGRFTISRDNSSSSNIGNNYVSW Construct KNTLYLQMNSLRAEDSAVYYCAKD YQQLPGTAPKLL ID:GVGATPYYFDYWGQGTLVTVSSAS IYDNNKRPSGIP PBD000101TKGPSVFPLAPSSKSTSGGTAALGCL DRFSGSKSGTSA 322 VKDYFPEPVTVSWNSGALTSGVHTFTLGITGLQTGDE PAVLQSSGLYSLSSVVTVPSSSLGTQ ADYYCGTWDSSTYICNVNHKPSNTKVDKKVEPKSCD LSAYVFGTGTKV KTHTCPPCPAPEAAGGPSVFLFPPKPTVLGQPKAAPSV KDTLMISRTPEVTCVVVSVSHEDPE TLFPPSSEELQANVKFNWYVDGVEVHNAKTKPREEQ KATLVCLISDFY YNSTYRVVSVLTVLHQDWLNGKEYPGAVTVAWKAD KCKVSNKALPAPIEKTISKAKGQPR SSPVKAGVETTTEPQVYTLPPSREEMTKNQVSLTCLV PSKQSNNKYAAS KGFYPSDIAVEWESNGQPENNYKTTSYLSLTPEQWKS PPVLDSDGSFFLYSRLTVDKSRWQQ HRSYSCQVTHEGGNVFSCSVMHEALHNHYTQKSLSL STVEKTVAPTEC SPG s PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1374 QSVLTQPPSVSA 1375 PSMB898 922GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGSGSTYYADSVKGRFTISRDNSSSSNIGINYVSW Construct KNTLYLQMNSLRAEDSAVYYCAKD YQQLPGTAPKLL ID:GVGATPYYFDYWGQGTLVTVSSAS IYDNNKRPSGIP PBD000101TKGPSVFPLAPSSKSTSGGTAALGCL DRFSGSKSGTSA 322 VKDYFPEPVTVSWNSGALTSGVHTFTLGITGLQTGDE PAVLQSSGLYSLSSVVTVPSSSLGTQ ADYYCGTWDSSTYICNVNHKPSNTKVDKKVEPKSCD LSAVVFGGGTKL KTHTCPPCPAPEAAGGPSVFLFPPKPTVLGQPKAAPSV KDTLMISRTPEVTCVVVSVSHEDPE TLFPPSSEELQANVKFNWYVDGVEVHNAKTKPREEQ KATLVCLISDFY YNSTYRVVSVLTVLHQDWLNGKEYPGAVTVAWKAD KCKVSNKALPAPIEKTISKAKGQPR SSPVKAGVETTTEPQVYTLPPSREEMTKNQVSLTCLV PSKQSNNKYAAS KGFYPSDIAVEWESNGQPENNYKTTSYLSLTPEQWKS PPVLDSDGSFFLYSRLTVDKSRWQQ HRSYSCQVTHEGGNVFSCSVMHEALHNHYTQKSLSL STVEKTVAPTEC SPG s PS3BEVQLVESGGGLVQPGGSLRLSCTAS 1376 QSVLTQPPSVSA 1377 PSMB899 912GFIFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGYGAPYYADTVKGRFTISRDNSSSNIGNNYVSW Construct SKNTLYLQMNSLRAEDTAVYYCAK YQQLPGTAPKLL ID:DGVGATPYYFDDWGQGILVTVSSA IFDNNKRPSGIPD PBD000101STKGPSVFPLAPSSKSTSGGTAALGC RFSGSKSGTSAT 324 LVKDYFPEPVTVSWNSGALTSGVHLGITGLQTGDEA TFPAVLQSSGLYSLSSVVTVPSSSLG DYYCGTWDSSLTQTYICNVNHKPSNTKVDKKVEPKS SAYVFGTGTKVT CDKTHTCPPCPAPEAAGGPSVFLFPPVLGQPKAAPSVT KPKDTLMISRTPEVTCVVVSVSHED LFPPSSEELQANPEVKFNWYVDGVEVHNAKTKPREE KATLVCLISDFY QYNSTYRVVSVLTVLHQDWLNGKEPGAVTVAWKAD YKCKVSNKALPAPIEKTISKAKGQP SSPVKAGVETTTREPQVYTLPPSREEMTKNQVSLTCL PSKQSNNKYAAS VKGFYPSDIAVEWESNGQPENNYKSYLSLTPEQWKS TTPPVLDSDGSFFLYSRLTVDKSRW HRSYSCQVTHEGQQGNVFSCSVMHEALHNHYTQKSL STVEKTVAPTEC SLSPG s PS3BEVQLVESGGGLVKPGGSLRLSCAAS 1378 QSVLTQPPSVSG 1379 PSMB889 930GFTFSRYNMNWVRQAPGKGLEWV APGQRVTISCTG HC2 SSINSNSRYIYYADSVKGRFTISRDSSSFNLGAGYDV Construct AKNSLYLQMNSLRAEDTAVYYCAK HWYQQVPGTVP ID:TMGDYYYYYGMDVWGQGTTVTVS KLLIYDNSNRPS PBD000101SASTKGPSVFPLAPSSKSTSGGTAAL GVPDRFSGSKSG 312 GCLVKDYFPEPVTVSWNSGALTSGTSASLAITGLQA VHTFPAVLQSSGLYSLSSVVTVPSSS EDETVYYCQSYLGTQTYICNVNHKPSNTKVDKKVEP DSSLSGVVFGGG KSCDKTHTCPPCPAPEAAGGPSVFLTKLTVLGQPKA FPPKPKDTLMISRTPEVTCVVVSVSH APSVTLFPPSSEEEDPEVKFNWYVDGVEVHNAKTKPR LQANKATLVCLI EEQYNSTYRVVSVLTVLHQDWLNGSDFYPGAVTVA KEYKCKVSNKALPAPIEKTISKAKG WKADSSPVKAGQPREPQVYTLPPSREEMTKNQVSLT VETTTPSKQSNN CLVKGFYPSDIAVEWESNGQPENNYKYAASSYLSLTP KTTPPVLDSDGSFFLYSRLTVDKSR EQWKSHRSYSCWQQGNVFSCSVMHEALHNHYTQK QVTHEGSTVEKT SLSLSPG VAPTECS PS3BEVQLVESGGGLVKPGGSLRLSCAAS 1380 SSELTQPPSVSG 1381 PSMB890 931GFTFSRYNMNWVRQAPGKGLEWV APGQRVTISCAG HC2 SSINSNSRYIYYADSVKGRFTISRDSSLSNIGAGYDVH Construct AKNSLYLQMNSLRAEDTAVYYCAK WYQQLPGTAPK ID:TMGDYYYYYGMDVWGQGTTVTVS LLIYGNINRLSG PBD000101SASTKGPSVFPLAPSSKSTSGGTAAL VPERFSGSKSGT 312 GCLVKDYFPEPVTVSWNSGALTSGSASLAITGLQAE VHTFPAVLQSSGLYSLSSVVTVPSSS DGADYYCQSYDLGTQTYICNVNHKPSNTKVDKKVEP SSLSSYVFGTGT KSCDKTHTCPPCPAPEAAGGPSVFLKVTVLGQPKAA FPPKPKDTLMISRTPEVTCVVVSVSH PSVTLFPPSSEELEDPEVKFNWYVDGVEVHNAKTKPR QANKATLVCLIS EEQYNSTYRVVSVLTVLHQDWLNGDFYPGAVTVAW KEYKCKVSNKALPAPIEKTISKAKG KADSSPVKAGVQPREPQVYTLPPSREEMTKNQVSLT ETTTPSKQSNNK CLVKGFYPSDIAVEWESNGQPENNYYAASSYLSLTPE KTTPPVLDSDGSFFLYSRLTVDKSR QWKSHRSYSCQWQQGNVFSCSVMHEALHNHYTQK VTHEGSTVEKTV SLSLSPG APTECS PS3BEVQLVESGGGVVQPGRSLRLSCAAS 1382 QSVLTQPPSASG 1383 PSMB891 926GFTFITYGMHWVRQAPGKGLEWVA TPGQGVTISCSG HC2 VVSFDESNKYYADSVKGRFTISRDNSSSNIGSNTVNW Construct SKNTLYLQMNSLRAEDTAVYYCAR FQQLPGTAPKLL ID:ALRDGNNWDYFNGMDVWGQGTT IYSDNQRPSGVP PBD000101 VTVSSASTKGPSVFPLAPSSKSTSGGDRFSGSKSGTSA 316 TAALGCLVKDYFPEPVTVSWNSGA SLAISGLQSEDELTSGVHTFPAVLQSSGLYSLSSVVT ADYYCAAWDDS VPSSSLGTQTYICNVNHKPSNTKVDLNGYVFGTGTK KKVEPKSCDKTHTCPPCPAPEAAGG VTVLGQPKAAPSPSVFLFPPKPKDTLMISRTPEVTCVV VTLFPPSSEELQA VSVSHEDPEVKFNWYVDGVEVHNANKATLVCLISDF KTKPREEQYNSTYRVVSVLTVLHQ YPGAVTVAWKADWLNGKEYKCKVSNKALPAPIEKTI DSSPVKAGVETT SKAKGQPREPQVYTLPPSREEMTKNTPSKQSNNKYA QVSLTCLVKGFYPSDIAVEWESNGQ ASSYLSLTPEQWPENNYKTTPPVLDSDGSFFLYSRLT KSHRSYSCQVTH VDKSRWQQGNVFSCSVMHEALHNEGSTVEKTVAPT HYTQKSLSLSPG ECS PS3B EVQLVESGGGVVQPGRSLRLSCAAS 1384QSVLTQPPSVSG 1385 PSMB892 928 GFTFITYGMHWVRQAPGKGLEWVA APGQRVTISCTG HC2VVSFDESNKYYADSVKGRFTISRDN SSSNIGADYDVH ConstructSKNTLYLQMNSLRAEDTAVYYCAR WYQHLPGTAPK ID: ALRDGNNWDYFNGMDVWGQGTTLLIYGNSNRPSG PBD000101 VTVSSASTKGPSVFPLAPSSKSTSGG VPDRFSGSKSGT 316TAALGCLVKDYFPEPVTVSWNSGA SASLAITGLQAE LTSGVHTFPAVLQSSGLYSLSSVVTDETDYYCQSYD VPSSSLGTQTYICNVNHKPSNTKVD SSLSGWVFGGGTKKVEPKSCDKTHTCPPCPAPEAAGG KLTVLGQPKAAP PSVFLFPPKPKDTLMISRTPEVTCVVSVTLFPPSSEELQ VSVSHEDPEVKFNWYVDGVEVHNA ANKATLVCLISDKTKPREEQYNSTYRVVSVLTVLHQ FYPGAVTVAWK DWLNGKEYKCKVSNKALPAPIEKTIADSSPVKAGVET SKAKGQPREPQVYTLPPSREEMTKN TTPSKQSNNKYAQVSLTCLVKGFYPSDIAVEWESNGQ ASSYLSLTPEQW PENNYKTTPPVLDSDGSFFLYSRLTKSHRSYSCQVTH VDKSRWQQGNVFSCSVMHEALHN EGSTVEKTVAPT HYTQKSLSLSPG ECS PS3BQVQLVESGGGVVQPGRSLRLSCVA 1386 QSVLTQPPSASG 1387 PSMB893 927SGFTFSSYGIHWVRQAPGKGLEWV TPGQGVTISCSG HC2 AVIWYDGSNKYYADSVKGRFTISRSSSNIGSNTVNW Construct DNSKNTLYLQMNSLRAEDTAVYYS FQQLPGTAPKLL ID:VRGVGPTSYYYNYGMDVWGQGTT IYSDNQRPSGVP PBD000101VTVSSASTKGPSVFPLAPSSKSTSGG DRFSGSKSGTSA 318 TAALGCLVKDYFPEPVTVSWNSGASLAISGLQSEDE LTSGVHTFPAVLQSSGLYSLSSVVT ADYYCAAWDDSVPSSSLGTQTYICNVNHKPSNTKVD LNGYVFGTGTK KKVEPKSCDKTHTCPPCPAPEAAGGVTVLGQPKAAPS PSVFLFPPKPKDTLMISRTPEVTCVV VTLFPPSSEELQAVSVSHEDPEVKFNWYVDGVEVHNA NKATLVCLISDF KTKPREEQYNSTYRVVSVLTVLHQYPGAVTVAWKA DWLNGKEYKCKVSNKALPAPIEKTI DSSPVKAGVETTSKAKGQPREPQVYTLPPSREEMTKN TPSKQSNNKYA QVSLTCLVKGFYPSDIAVEWESNGQASSYLSLTPEQW PENNYKTTPPVLDSDGSFFLYSRLT KSHRSYSCQVTHVDKSRWQQGNVFSCSVMHEALHN EGSTVEKTVAPT HYTQKSLSLSPG ECS PS3BQVQLVESGGGVVQPGRSLRLSCVA 1388 QSVLTQPPSVSG 1389 PSMB894 929SGFTFSSYGIHWVRQAPGKGLEWV APGQRVTISCTG HC2 AVIWYDGSNKYYADSVKGRFTISRSSSNIGADYDVH Construct DNSKNTLYLQMNSLRAEDTAVYYS WYQHLPGTAPK ID:VRGVGPTSYYYNYGMDVWGQGTT LLIYGNSNRPSG PBD000101VTVSSASTKGPSVFPLAPSSKSTSGG VPDRFSGSKSGT 318 TAALGCLVKDYFPEPVTVSWNSGASASLAITGLQAE LTSGVHTFPAVLQSSGLYSLSSVVT DETDYYCQSYDVPSSSLGTQTYICNVNHKPSNTKVD SSLSGWVFGGGT KKVEPKSCDKTHTCPPCPAPEAAGGKLTVLGQPKAAP PSVFLFPPKPKDTLMISRTPEVTCVV SVTLFPPSSEELQVSVSHEDPEVKFNWYVDGVEVHNA ANKATLVCLISD KTKPREEQYNSTYRVVSVLTVLHQFYPGAVTVAWK DWLNGKEYKCKVSNKALPAPIEKTI ADSSPVKAGVETSKAKGQPREPQVYTLPPSREEMTKN TTPSKQSNNKYA QVSLTCLVKGFYPSDIAVEWESNGQASSYLSLTPEQW PENNYKTTPPVLDSDGSFFLYSRLT KSHRSYSCQVTHVDKSRWQQGNVFSCSVMHEALHN EGSTVEKTVAPT HYTQKSLSLSPG ECS PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1390 QSVLTQPPSVSA 1391 PSMB895 932GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGIGSTYYADSVKGRFTISRDNSSSSNIGNNYVSW Construct KNTLWLQMNSLRAEDTAVYYCAK YQQLPGTAPKLL ID:DGVGATPYYFDYWGQGTLVTVSSA IYDNNKRPSGIP PBD000101STKGPSVFPLAPSSKSTSGGTAALGC DRFSGSKSGTSA 320 LVKDYFPEPVTVSWNSGALTSGVHTLGITGLQTGDE TFPAVLQSSGLYSLSSVVTVPSSSLG ADYYCGTWDSSTQTYICNVNHKPSNTKVDKKVEPKS LSAYVFGTGTKV CDKTHTCPPCPAPEAAGGPSVFLFPPTVLGQPKAAPSV KPKDTLMISRTPEVTCVVVSVSHED TLFPPSSEELQANPEVKFNWYVDGVEVHNAKTKPREE KATLVCLISDFY QYNSTYRVVSVLTVLHQDWLNGKEPGAVTVAWKAD YKCKVSNKALPAPIEKTISKAKGQP SSPVKAGVETTTREPQVYTLPPSREEMTKNQVSLTCL PSKQSNNKYAAS VKGFYPSDIAVEWESNGQPENNYKSYLSLTPEQWKS TTPPVLDSDGSFFLYSRLTVDKSRW HRSYSCQVTHEGQQGNVFSCSVMHEALHNHYTQKSL STVEKTVAPTEC SLSPG s PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1392 QSVLTQPPSVSA 1393 PSMB896 934GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGIGSTYYADSVKGRFTISRDNSSSSNIGINYVSW Construct KNTLWLQMNSLRAEDTAVYYCAK YQQLPGTAPKLL ID:DGVGATPYYFDYWGQGTLVTVSSA IYDNNKRPSGIP PBD000101STKGPSVFPLAPSSKSTSGGTAALGC DRFSGSKSGTSA 320 LVKDYFPEPVTVSWNSGALTSGVHTLGITGLQTGDE TFPAVLQSSGLYSLSSVVTVPSSSLG ADYYCGTWDSSTQTYICNVNHKPSNTKVDKKVEPKS LSAVVFGGGTKL CDKTHTCPPCPAPEAAGGPSVFLFPPTVLGQPKAAPSV KPKDTLMISRTPEVTCVVVSVSHED TLFPPSSEELQANPEVKFNWYVDGVEVHNAKTKPREE KATLVCLISDFY QYNSTYRVVSVLTVLHQDWLNGKEPGAVTVAWKAD YKCKVSNKALPAPIEKTISKAKGQP SSPVKAGVETTTREPQVYTLPPSREEMTKNQVSLTCL PSKQSNNKYAAS VKGFYPSDIAVEWESNGQPENNYKSYLSLTPEQWKS TTPPVLDSDGSFFLYSRLTVDKSRW HRSYSCQVTHEGQQGNVFSCSVMHEALHNHYTQKSL STVEKTVAPTEC SLSPG s PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1394 QSVLTQPPSVSA 1395 PSMB897 933GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGSGSTYYADSVKGRFTISRDNSSSSNIGNNYVSW Construct KNTLYLQMNSLRAEDSAVYYCAKD YQQLPGTAPKLL ID:GVGATPYYFDYWGQGTLVTVSSAS IYDNNKRPSGIP PBD000101TKGPSVFPLAPSSKSTSGGTAALGCL DRFSGSKSGTSA 322 VKDYFPEPVTVSWNSGALTSGVHTFTLGITGLQTGDE PAVLQSSGLYSLSSVVTVPSSSLGTQ ADYYCGTWDSSTYICNVNHKPSNTKVDKKVEPKSCD LSAYVFGTGTKV KTHTCPPCPAPEAAGGPSVFLFPPKPTVLGQPKAAPSV KDTLMISRTPEVTCVVVSVSHEDPE TLFPPSSEELQANVKFNWYVDGVEVHNAKTKPREEQ KATLVCLISDFY YNSTYRVVSVLTVLHQDWLNGKEYPGAVTVAWKAD KCKVSNKALPAPIEKTISKAKGQPR SSPVKAGVETTTEPQVYTLPPSREEMTKNQVSLTCLV PSKQSNNKYAAS KGFYPSDIAVEWESNGQPENNYKTTSYLSLTPEQWKS PPVLDSDGSFFLYSRLTVDKSRWQQ HRSYSCQVTHEGGNVFSCSVMHEALHNHYTQKSLSL STVEKTVAPTEC SPG S PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1396 QSVLTQPPSVSA 1397 PSMB898 935GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGSGSTYYADSVKGRFTISRDNSSSSNIGINYVSW Construct KNTLYLQMNSLRAEDSAVYYCAKD YQQLPGTAPKLL ID:GVGATPYYFDYWGQGTLVTVSSAS IYDNNKRPSGIP PBD000101TKGPSVFPLAPSSKSTSGGTAALGCL DRFSGSKSGTSA 322 VKDYFPEPVTVSWNSGALTSGVHTFTLGITGLQTGDE PAVLQSSGLYSLSSVVTVPSSSLGTQ ADYYCGTWDSSTYICNVNHKPSNTKVDKKVEPKSCD LSAVVFGGGTKL KTHTCPPCPAPEAAGGPSVFLFPPKPTVLGQPKAAPSV KDTLMISRTPEVTCVVVSVSHEDPE TLFPPSSEELQANVKFNWYVDGVEVHNAKTKPREEQ KATLVCLISDFY YNSTYRVVSVLTVLHQDWLNGKEYPGAVTVAWKAD KCKVSNKALPAPIEKTISKAKGQPR SSPVKAGVETTTEPQVYTLPPSREEMTKNQVSLTCLV PSKQSNNKYAAS KGFYPSDIAVEWESNGQPENNYKTTSYLSLTPEQWKS PPVLDSDGSFFLYSRLTVDKSRWQQ HRSYSCQVTHEGGNVFSCSVMHEALHNHYTQKSLSL STVEKTVAPTEC SPG s PS3BEVQLVESGGGLVQPGGSLRLSCTAS 1398 QSVLTQPPSVSA 1399 PSMB899 925GFIFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGYGAPYYADTVKGRFTISRDNSSSNIGNNYVSW Construct SKNTLYLQMNSLRAEDTAVYYCAK YQQLPGTAPKLL ID:DGVGATPYYFDDWGQGILVTVSSA IFDNNKRPSGIPD PBD000101STKGPSVFPLAPSSKSTSGGTAALGC RFSGSKSGTSAT 324 LVKDYFPEPVTVSWNSGALTSGVHLGITGLQTGDEA TFPAVLQSSGLYSLSSVVTVPSSSLG DYYCGTWDSSLTQTYICNVNHKPSNTKVDKKVEPKS SAYVFGTGTKVT CDKTHTCPPCPAPEAAGGPSVFLFPPVLGQPKAAPSVT KPKDTLMISRTPEVTCVVVSVSHED LFPPSSEELQANPEVKFNWYVDGVEVHNAKTKPREE KATLVCLISDFY QYNSTYRVVSVLTVLHQDWLNGKEPGAVTVAWKAD YKCKVSNKALPAPIEKTISKAKGQP SSPVKAGVETTTREPQVYTLPPSREEMTKNQVSLTCL PSKQSNNKYAAS VKGFYPSDIAVEWESNGQPENNYKSYLSLTPEQWKS TTPPVLDSDGSFFLYSRLTVDKSRW HRSYSCQVTHEGQQGNVFSCSVMHEALHNHYTQKSL STVEKTVAPTEC SLSPG S PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1400 QSVLTQPPSVSA 1401 PSMB946 1352GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGIGSTYYADSVKGRFTISRDNSSSSNIGNNYVSW Construct KNTLWLQMNSLRAEDTAVYYCAK YQQLPGTAPKLL ID:DGVGATPYYFDYWGQGTLVTVSSA IYDNNKRPSGIP PBD000108STKGPSVFPLAPSSKSTSGGTAALGC DRFSGSKSGTSA 502 LVKDYFPEPVTVSWNSGALTSGVHTLGITGLQTGDE TFPAVLQSSGLYSLSSVVTVPSSSLG ADYYCGTWDSSTQTYICNVNHKPSNTKVDKKVEPKS LSAYVFGTGTKV CDKTHTCPPCPAPEAAGGPSVFLFPPTVLGQPKAAPSV KPKDTLMISRTPEVTCVVVSVSHED TLFPPSSEELQANPEVKFNWYVDGVEVHNAKTKPREE KATLVCLISDFY QYNSTYRVVSVLTVLHQDWLNGKEPGAVTVAWKAD YKCKVSNKALPAPIEKTISKAKGQP SSPVKAGVETTTREPQVYTLPPSREEMTKNQVSLTCL PSKQSNNKYAAS VKGFYPSDIAVEWESNGQPENNYKSYLSLTPEQWKS TTPPVLDSDGSFFLYSRLTVDKSRW HRSYSCQVTHEGQQGNVFSCSVMHEALHNHYTQKSL STVEKTVAPTEC SLSPGK s PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1402 QSVLTQPPSVSA 1403 PSMB847 1353GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGIGSTYYADSVKGRFTISRDNSSSSNIGINYVSW Construct KNTLWLQMNSLRAEDTAVYYCAK YQQLPGTAPKLL ID:DGVGATPYYFDYWGQGTLVTVSSA IYDNNKRPSGIP PBD000108STKGPSVFPLAPSSKSTSGGTAALGC DRFSGSKSGTSA 503 LVKDYFPEPVTVSWNSGALTSGVHTLGITGLQTGDE TFPAVLQSSGLYSLSSVVTVPSSSLG ADYYCGTWDSSTQTYICNVNHKPSNTKVDKKVEPKS LSAVVFGGGTKL CDKTHTCPPCPAPEAAGGPSVFLFPPTVLGQPKAAPSV KPKDTLMISRTPEVTCVVVSVSHED TLFPPSSEELQANPEVKFNWYVDGVEVHNAKTKPREE KATLVCLISDFY QYNSTYRVVSVLTVLHQDWLNGKEPGAVTVAWKAD YKCKVSNKALPAPIEKTISKAKGQP SSPVKAGVETTTREPQVYTLPPSREEMTKNQVSLTCL PSKQSNNKYAAS VKGFYPSDIAVEWESNGQPENNYKSYLSLTPEQWKS TTPPVLDSDGSFFLYSRLTVDKSRW HRSYSCQVTHEGQQGNVFSCSVMHEALHNHYTQKSL STVEKTVAPTEC SLSPGK s PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1404 QSVLTQPPSVSA 1405 PSMB848 1354GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGSGSTYYADSVKGRFTISRDNSSSSNIGNNYVSW Construct KNTLYLQMNSLRAEDSAVYYCAKD YQQLPGTAPKLL ID:GVGATPYYFDYWGQGTLVTVSSAS IYDNNKRPSGIP PBD000108TKGPSVFPLAPSSKSTSGGTAALGCL DRFSGSKSGTSA 504 VKDYFPEPVTVSWNSGALTSGVHTFTLGITGLQTGDE PAVLQSSGLYSLSSVVTVPSSSLGTQ ADYYCGTWDSSTYICNVNHKPSNTKVDKKVEPKSCD LSAYVFGTGTKV KTHTCPPCPAPEAAGGPSVFLFPPKPTVLGQPKAAPSV KDTLMISRTPEVTCVVVSVSHEDPE TLFPPSSEELQANVKFNWYVDGVEVHNAKTKPREEQ KATLVCLISDFY YNSTYRVVSVLTVLHQDWLNGKEYPGAVTVAWKAD KCKVSNKALPAPIEKTISKAKGQPR SSPVKAGVETTTEPQVYTLPPSREEMTKNQVSLTCLV PSKQSNNKYAAS KGFYPSDIAVEWESNGQPENNYKTTSYLSLTPEQWKS PPVLDSDGSFFLYSRLTVDKSRWQQ HRSYSCQVTHEGGNVFSCSVMHEALHNHYTQKSLSL STVEKTVAPTEC SPGK S PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1406 QSVLTQPPSVSA 1407 PSMB849 1355GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGSGSTYYADSVKGRFTISRDNSSSSNIGINYVSW Construct KNTLYLQMNSLRAEDSAVYYCAKD YQQLPGTAPKLL ID:GVGATPYYFDYWGQGTLVTVSSAS IYDNNKRPSGIP PBD000108TKGPSVFPLAPSSKSTSGGTAALGCL DRFSGSKSGTSA 505 VKDYFPEPVTVSWNSGALTSGVHTFTLGITGLQTGDE PAVLQSSGLYSLSSVVTVPSSSLGTQ ADYYCGTWDSSTYICNVNHKPSNTKVDKKVEPKSCD LSAVVFGGGTKL KTHTCPPCPAPEAAGGPSVFLFPPKPTVLGQPKAAPSV KDTLMISRTPEVTCVVVSVSHEDPE TLFPPSSEELQANVKFNWYVDGVEVHNAKTKPREEQ KATLVCLISDFY YNSTYRVVSVLTVLHQDWLNGKEYPGAVTVAWKAD KCKVSNKALPAPIEKTISKAKGQPR SSPVKAGVETTTEPQVYTLPPSREEMTKNQVSLTCLV PSKQSNNKYAAS KGFYPSDIAVEWESNGQPENNYKTTSYLSLTPEQWKS PPVLDSDGSFFLYSRLTVDKSRWQQ HRSYSCQVTHEGGNVFSCSVMHEALHNHYTQKSLSL STVEKTVAPTEC SPGK s PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1408 QSVLTQPPSVSA 1409 PSMB946 1356GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGIGSTYYADSVKGRFTISRDNSSSSNIGNNYVSW Construct KNTLWLQMNSLRAEDTAVYYCAK YQQLPGTAPKLL ID:DGVGATPYYFDYWGQGTLVTVSSA IYDNNKRPSGIP PBD000108STKGPSVFPLAPSSKSTSGGTAALGC DRFSGSKSGTSA 502 LVKDYFPEPVTVSWNSGALTSGVHTLGITGLQTGDE TFPAVLQSSGLYSLSSVVTVPSSSLG ADYYCGTWDSSTQTYICNVNHKPSNTKVDKKVEPKS LSAYVFGTGTKV CDKTHTCPPCPAPEAAGGPSVFLFPPTVLGQPKAAPSV KPKDTLMISRTPEVTCVVVSVSHED TLFPPSSEELQANPEVKFNWYVDGVEVHNAKTKPREE KATLVCLISDFY QYNSTYRVVSVLTVLHQDWLNGKEPGAVTVAWKAD YKCKVSNKALPAPIEKTISKAKGQP SSPVKAGVETTTREPQVYTLPPSREEMTKNQVSLTCL PSKQSNNKYAAS VKGFYPSDIAVEWESNGQPENNYKSYLSLTPEQWKS TTPPVLDSDGSFFLYSRLTVDKSRW HRSYSCQVTHEGQQGNVFSCSVMHEALHNHYTQKSL STVEKTVAPTEC SLSPGK S PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1410 QSVLTQPPSVSA 1411 PSMB847 1357GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGIGSTYYADSVKGRFTISRDNSSSSNIGINYVSW Construct KNTLWLQMNSLRAEDTAVYYCAK YQQLPGTAPKLL ID:DGVGATPYYFDYWGQGTLVTVSSA IYDNNKRPSGIP PBD000108STKGPSVFPLAPSSKSTSGGTAALGC DRFSGSKSGTSA 503 LVKDYFPEPVTVSWNSGALTSGVHTLGITGLQTGDE TFPAVLQSSGLYSLSSVVTVPSSSLG ADYYCGTWDSSTQTYICNVNHKPSNTKVDKKVEPKS LSAVVFGGGTKL CDKTHTCPPCPAPEAAGGPSVFLFPPTVLGQPKAAPSV KPKDTLMISRTPEVTCVVVSVSHED TLFPPSSEELQANPEVKFNWYVDGVEVHNAKTKPREE KATLVCLISDFY QYNSTYRVVSVLTVLHQDWLNGKEPGAVTVAWKAD YKCKVSNKALPAPIEKTISKAKGQP SSPVKAGVETTTREPQVYTLPPSREEMTKNQVSLTCL PSKQSNNKYAAS VKGFYPSDIAVEWESNGQPENNYKSYLSLTPEQWKS TTPPVLDSDGSFFLYSRLTVDKSRW HRSYSCQVTHEGQQGNVFSCSVMHEALHNHYTQKSL STVEKTVAPTEC SLSPGK s PS3BEVQLVESGGGLVQPGGSLRLSCAAS 1412 QSVLTQPPSVSA 1413 PSMB849 1358GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGSGSTYYADSVKGRFTISRDNSSSSNIGINYVSW Construct KNTLYLQMNSLRAEDSAVYYCAKD YQQLPGTAPKLL ID:GVGATPYYFDYWGQGTLVTVSSAS IYDNNKRPSGIP PBD000108TKGPSVFPLAPSSKSTSGGTAALGCL DRFSGSKSGTSA 505 VKDYFPEPVTVSWNSGALTSGVHTFTLGITGLQTGDE PAVLQSSGLYSLSSVVTVPSSSLGTQ ADYYCGTWDSSTYICNVNHKPSNTKVDKKVEPKSCD LSAVVFGGGTKL KTHTCPPCPAPEAAGGPSVFLFPPKPTVLGQPKAAPSV KDTLMISRTPEVTCVVVSVSHEDPE TLFPPSSEELQANVKFNWYVDGVEVHNAKTKPREEQ KATLVCLISDFY YNSTYRVVSVLTVLHQDWLNGKEYPGAVTVAWKAD KCKVSNKALPAPIEKTISKAKGQPR SSPVKAGVETTTEPQVYTLPPSREEMTKNQVSLTCLV PSKQSNNKYAAS KGFYPSDIAVEWESNGQPENNYKTTSYLSLTPEQWKS PPVLDSDGSFFLYSRLTVDKSRWQQ HRSYSCQVTHEGGNVFSCSVMHEALHNHYTQKSLSL STVEKTVAPTEC SPGK S PSMBEVQLVESGGGLVQPGGSLRLSCAAS 1414 QSVLTQPPSVSA 1415 PSMB848 937GFTFSSYAMSWVRQAPGKGLEWVS APGQKVTISCSG HC2 AISGGSGSTYYADSVKGRFTISRDNSSSSNIGNNYVSW Construct KNTLYLQMNSLRAEDSAVYYCAKD YQQLPGTAPKLL ID:GVGATPYYFDYWGQGTLVTVSSAS IYDNNKRPSGIP PBD000101TKGPSVFPLAPSSKSTSGGTAALGCL DRFSGSKSGTSA 322 VKDYFPEPVTVSWNSGALTSGVHTFTLGITGLQTGDE PAVLQSSGLYSLSSVVTVPSSSLGTQ ADYYCGTWDSSTYICNVNHKPSNTKVDKKVEPKSCD LSAYVFGTGTKV KTHTCPPCPAPEAAGGPSVFLFPPKPTVLGQPKAAPSV KDTLMISRTPEVTCVVVSVSHEDPE TLFPPSSEELQANVKFNWYVDGVEVHNAKTKPREEQ KATLVCLISDFY YNSTYRVVSVLTVLHQDWLNGKEYPGAVTVAWKAD KCKVSNKALPAPIEKTISKAKGQPR SSPVKAGVETTTEPQVYTLPPSREEMTKNQVSLTCLV PSKQSNNKYAAS KGFYPSDIAVEWESNGQPENNYKTTSYLSLTPEQWKS PPVLDSDGSFFLYSRLTVDKSRWQQ HRSYSCQVTHEGGNVFSCSVMHEALHNHYTQKSLSL STVEKTVAPTEC SPG S

Example 8.2: Analytical Characterization of Bispecific Anti-PSMAxCD3Antibodies

The protein concentration for each purified bispecific Ab was determinedby measuring the absorbance at 280 nm on a NANODROP1000spectrophotometer or Trinean DROPSENSE96 multichannel spectrophotometerand calculated using the extinction coefficient based on the amino acidsequence. SE HPLC of the purified antibodies was performed by runningsamples on a TOSOH TSKgel BioAssist G3SWxl column, in 0.2 M Na PhosphatepH 6.8 at 1 mL/min on a Waters Alliance HPLC for 20 min. The columneffluent was monitored by absorbance at 280 nm. Anti-PSMA-CD3 bispecificantibodies were further analyzed by Intact Mass Analysis to determineappropriate formation of heterodimers.

Example 9: Epitope Mapping of Anti-PSMAxCD3 Antibodies Example 9.1:HDX-MS Epitope Mapping

The epitope of two anti-PSMA/CD3 bispecific antibodies PS3B1352 andPS3B1353 were determined by hydrogen-deuterium exchange massspectrometry (HDX-MS). See FIG. 14 . Human PSMA antigen was used forepitope mapping experiment. See FIG. 15 .

On-Exchange Experiment for HDX-MS. Briefly, on-exchange reaction wasinitiated by mixing 10 μL of 6.0 μM human PSMA with or without 7.3 μMantibody and 30 μL of H₂O or a deuterated buffer (20 mM MES, pH 6.4, 150mM NaCl in 95% D₂O or 20 mM Tris, pH 8.4, 150 mM NaCl in 95% D₂O). Thereaction mixture was incubated for 15, 50, 150, 500, or 1,500 s at 23°C. and quenched at the different time points described by the additionof chilled 40 μL of 8 M urea, 1 M TCEP, pH 3.0. The quenched solutionswere analyzed immediately.

General Procedure for HDX-MS Data Acquisition. HDX-MS sample preparationwas performed with automated HDx system (LEAP Technologies, Morrisville,N.C.). The columns and pump were; protease, protease type XIII (proteasefrom Aspergillus saitoi, type XIII)/pepsin column (w/w, 1:1; 2.1×30 mm)(NovaBioAssays Inc., Woburn, Mass.); trap, ACQUITY UPLC BEH C18 VanGuardPre-column (2.1×5 mm) (Waters, Milford, Mass.), analytical, Accucore C18(2.1×100 mm) (Thermo Fisher Scientific, Waltham, Mass.); and LC pump,VH-P10-A (Thermo Fisher Scientific). The loading pump (from the proteasecolumn to the trap column) was set at 600 μL/min with 99% water, 1%acetonitrile, 0.1% formic acid. The gradient pump (from the trap columnto the analytical column) was set from 8% to 28% acetonitrile in 0.1%aqueous formic acid in 20 min at 100 μL/min.

MS Data Acquisition. Mass spectrometric analyses were carried out usingan LTQ™ Orbitrap Fusion Lumos mass spectrometer (Thermo FisherScientific) with the capillary temperature at 275° C., resolution150,000, and mass range (m/z) 300-2,000.

HDX-MS Data Extraction. BioPharma Finder 2.0 (Thermo Fisher Scientific)was used for the peptide identification of non-deuterated samples priorto the HDX experiments. HDExaminer version 2.4 (Sierra Analytics,Modesto, Calif.) was used to extract centroid values from the MS rawdata files for the HDX experiments.

Example 10: Characterization of Bispecific Anti-PSMAxCD3 AntibodiesExample 10.1: Binding Affinity of Bispecific Anti-PSMAxCD3 Antibodies toHuman PSMA

The binding affinity of anti-PSMA to the recombinant human, cynomolgusor mouse PSMA was determined by surface plasmon resonance (SPR) using aBiacore 8K instrument\. The antibodies were captured on a goat anti-Fcantibody-modified C1 chip and titrated with 3-fold serial dilutions ofPSMA antigen spanning concentrations of 100 nM to 11.1 nM. Theassociation and dissociation were monitored for 3 and 15 minutes,respectively, using a flow rate of 50 μL/min. Raw binding data wasreferenced by subtracting the analyte binding signals from blanks andanalyzed using a 1:1 Langmuir binding model using the Biacore Insightevaluation software to obtain the kinetics which were used to calculatethe binding affinity. Kd data are summarized in Table 64. The anti-PSMAwere captured using an anti-human Fc antibody and the antigens wereinjected in solution.

TABLE 64 Affinities (KD) for the interaction of anti-PSMA × CD3bispecific antibodies with human PSMA as obtained by the BIACORE (SPR)method. Name KD (M) PS3B917 ≤2.33E−11   PS3B918 poor fit PS3B913≤2.40E−10   PS3B915 1.89E−09 PS3B914 Low/No binding PS3B916 1.75E−09PS3B919 8.23E−10 PS3B921 ≤8.64E−11   PS3B920 1.35E−09 PS3B922 4.10E−10PS3B912 1.74E−10 PS3B930 ≤2.26E−11   PS3B931 poor fit PS3B926≤2.53E−10   PS3B928 2.37E−09 PS3B927 1.11E−09 PS3B929 1.83E−09 PS3B9327.87E−10 PS3B934 ≤8.73E−11   PS3B933 1.44E−09 PS3B935 4.54E−10 PS3B9251.69E−10

The binding affinity of anti-PSMA antibodies to the recombinant humanPSMA was determined by surface plasmon resonance (SPR) using a BIACORE8K instrument (ELN PSMA-00702). The antibodies were captured on a goatanti-Fc antibody-modified C1 chip and titrated with 3-fold serialdilutions of PSMA antigen spanning concentrations of 1 nM to 100 nMhuman PSMA. The association and dissociation were monitored for 3 and 30minutes, respectively, using a flow rate of 50 μL/min. Raw binding datawas referenced by subtracting the analyte binding signals from blanksand analyzed using a 1:1 Langmuir binding model by the Biacore Insightevaluation software to obtain the kinetics which were used to calculatethe binding affinity. The kinetic parameter of binding of selectedantibodies are shown in Table 65. The anti-PSMA were captured using ananti-human Fc antibody and the antigens were injected in solution.

TABLE 65 Affinities (KD) for the interaction of anti-PSMA antibodieswith Human PSMA as obtained by the Biacore (SPR) method. ka (1/Ms) kd(1/s) KD (M) PSMB1045 2.09E+05 6.98E−05 3.33E−10 PSMB1049 1.07E+057.02E−05 6.57E−10 PSMB1051 7.71E+04 7.75E−05 1.01E−09 PSMB1041 2.07E+054.35E−04 2.11E−09 PSMB1068 1.51E+05 1.68E−04 1.11E−09 PSMB1052 1.96E+053.15E−04 1.61E−09 PSMB1069 1.39E+05 1.97E−04 1.42E−09 PSMB1047 6.94E+051.75E−03 2.52E−09 PSMB1075 5.26E+05 4.92E−04 9.36E−10 PSMB1060 1.17E+051.19E−04 1.02E−09

The binding affinity of anti-PSMA to the recombinant human or cynomolgusPSMA was determined by surface plasmon resonance (SPR) using a Biacore8K instrument (ELN PSMA-00721). The antibodies were captured on a goatanti-Fc antibody-modified C1 chip and titrated with 3-fold serialdilutions of PSMA antigen spanning concentrations of 100 nM to 3.7 nM(huma PSMA) or 100 nM to 3.7 nM, or 22.2-600 nM for cyno PSMA. Theassociation and dissociation were monitored for 3 and 60 minutes,respectively, using a flow rate of 50 μL/min. Raw binding data wasreferenced by subtracting the analyte binding signals from blanks andanalyzed using a 1:1 Langmuir binding model by the Biacore Insightevaluation software to obtain the kinetics which were used to calculatethe binding affinity. The kinetic parameter of binding of selectedantibodies are shown in Table 66. The anti-PSMA were captured using ananti-human Fc antibody and the antigens were injected in solution.

TABLE 66 Affinities (KD)* for the interaction of anti-PSMA antibodieswith Human, cyno or Mouse PSMA as obtained by the Biacore (SPR) method.Avg. ka 95% Avg. kd 95% Avg. KD 95% Protein name Antigen (1/Ms) CI (1/s)CI (M) CI PS3B1391.002 cy PSMA 1.57E+05 5.77E+04 1.96E−05 6.59E−061.26E−10 3.72E−11 PS3B1391.002 hu PSMA 4.80E+05 4.80E−05 1.00E−10PS3B1508.003 cy PSMA 1.46E+06 3.23E−03 2.20E−09 PS3B1508.003 hu PSMA2.32E+05 8.89E−05 3.82E−10 PS3B1396.002 cy PSMA 5.87E+05 1.90E−035.07E−09 PS3B1396.002 hu PSMA 2.24E+05 8.82E−05 3.95E−10 PS3B1505.001 cyPSMA 4.29E+05 1.24E−03 5.01E−09 PS3B1505.001 hu PSMA 2.32E+05 9.10E−053.93E−10 *Data in row 1 is an average of 3. Data in other rows anaverage of 2.

Example 10.2: Thermal Stability of Bispecific Anti-PSMAxCD3 Antibodies

The thermal stability (conformational stability information including,Tm and Tagg) of anti-PSMAxCD3 antibodies was determined by nanoDSFmethod using a Prometheus instrument as described above. Briefly,measurements were made by loading sample into 24 well capillary from a384 well sample plate. Duplicate runs were performed. The thermal scansspan from 20° C. to 95° C. at a rate of 1.0° C./minute. The data wasprocessed to obtain integrated data and first derivation analysis for330 nm, 350 nm, Ratio 330/350, and scatter data from which thermaltransitions, onset of unfolding, Tm and Tagg were obtained andsummarized in Table 67.

TABLE 67 Thermal stability data for bispecific anti-PSMA × CD3antibodies as obtained using a nanoDSF instrument. Name Tage T_(m)lT_(m)2 T_(m)3 PS3B917 68.2° C. 63.5° C. 68.8° C. PS3B918 67.8° C. 63.4°C. 68.2° C. PS3B913 75.1° C. 63.4° C. 76.3° C. PS3B915 69.2° C. 63.5° C.PS3B914 64.5° C. 61.9° C. 67.0° C. 75.8° C. PS3B916 59.1° C. 59.9° C.PS3B919 81.0° C. 63.5° C. 68.3° C. 85.7° C. PS3B921 78.9° C. 63.5° C.80.8° C. PS3B920 82.6° C. 63.4° C. 87.5° C. PS3B922 80.5° C. 63.3° C.83.4° C. PS3B912 77.1° C. 63.6° C. 78.4° C. PS3B930 72.2° C. 70.1° C.PS3B931 72.2° C. 69.5° C. PS3B926 75.6° C. 70.2° C. 75.6° C. PS3B92872.2° C. 69.1° C. PS3B927 69.1° C. 66.5° C. 69.4° C. PS3B929 69.1° C.58.0° C. 68.0° C. PS3B932 82.7° C. 70.4° C. 85.8° C. PS3B934 79.3° C.70.4° C. 80.9° C. PS3B933 83.7° C. 70.0° C. 87.8° C. PS3B935 81.5° C.70.2° C. 83.7° C. PS3B925 77.4° C. 70.3° C. 77.9° C.

Example 10.3: Binding of Bispecific PSMAxCD3 Antibodies on PSMA+ Cells

Selected bispecific PSMAxCD3 antibodies were assessed for their abilityto bind prostate cancer cell lines expressing PSMA.

22RV1 and C4-2B cells were plated at 50,000 cells per well in 50 μl ofassay medium (RPMI, 10% HI FBS) in V bottom plates. Serial dilutions ofantibodies were prepared in assay medium with 50 μl of antibodydilutions added to the plates containing cells. The plates wereincubated for 60 min at 37° C. at which time 100 μl of staining buffer(Becton Dickinson Cat #554657) was added to all wells of each plate. Theplates were centrifuged at 300×G for 5 minutes and the medium wasremoved from the wells. 200 μl of staining buffer was added to all wellsof each plate. The plates were centrifuged at 300×G for 5 minutes andthe medium was removed from the wells. 50 μl of 2 μg/mlAlexaFluor647-labeled goat anti-human Fc was added to all wells of theplates and the plates were incubated for 30 minutes at 4° C. 150 μl ofstaining buffer was added to all wells of each plate. The plates werecentrifuged at 300×G for 5 minutes and the medium was removed from thewells. Two hundred microliters of running buffer (Staining buffer plus 1mM EDTA, 0.1% pluronic acid was added to all wells of the plates. Theplates were centrifuged at 300×G for 5 minutes and the medium wasremoved from the wells. Thirty microliters running buffer was added toall wells with cells and the plates were analyzed on the IQUE Plusinstrument (Sartorius). Briefly, cells were gated on a FCS vs. SSC gateto eliminate cellular debris, then the cell populations were gated onsinglet cells. Antibody binding was assessed in the red laser channelSignal (Mab plus secondary antibody) to background (secondary antibodyonly) ratios were calculated for each plate and the resultant data wasplotted vs. bispecific antibody concentration in GeneData Screener using4 parameter curve fitting to generate EC50 values, summarized in Table68.

TABLE 68 EC50 values of bispecific PSMA × CD3 antibodies binding toPSMA-expressing cell lines in flow cytometry assays. Name 22RV1 bindingEC50 [M] C4-2B binding EC50 [M] PS3B917 7.50E−10 1.90E−09 PS3B9186.00E−08 6.00E−08 PS3B913 3.42E−09 4.56E−09 PS3B915 6.00E−08 6.00E−08PS3B914 6.00E−08 6.00E−08 PS3B916 6.00E−08 6.00E−08 PS3B919 2.49E−082.33E−08 PS3B921 4.03E−09 6.44E−09 PS3B920 6.00E−08 6.00E−08 PS3B9223.47E−08 4.41E−08 PS3B912 5.40E−08 3.68E−08 PS3B930 1.06E−09 3.01E−09PS3B931 6.00E−08 6.00E−08 PS3B926 2.90E−09 4.55E−09 PS3B928 6.00E−086.00E−08 PS3B927 6.00E−08 6.00E−08 PS3B929 5.48E−08 6.00E−08 PS3B9324.87E−08 6.00E−08 PS3B934 4.96E−09 1.10E−08 PS3B933 6.00E−08 6.00E−08PS3B935 3.95E−08 8.22E−09 PS3B925 2.12E−08 1.17E−08

Binding of anti-PSMA/CD3 bispecific on PAN-T cells via Flow. Human PAN-TCells (Biological Specialty Corporation, Colmar, Pa.) were thawed andtransferred to a 15 mL conical with DPBS. The cells were centrifuged1300 rpm for 5 minutes. DPBS was aspirated and cells were re-suspendedin DPBS. The cells were counted using the Vi-cell XR cell viabilityanalyzer and were plated at 100K/well in 100 uL DPBS. The plate wascentrifuged 1200 rpm for 3 minutes and washed 2× with DPBS. Cells werestained with Violet Live/Dead stain (Thermo-Fisher) and incubated at RTin the dark for 25 min. The cells were centrifuged and washed 2× withFACS staining buffer (BD Pharmingen). Test antibodies were diluted to afinal starting concentration of 1 μM in FACS staining buffer and 3-foldserial dilutions were prepared from the starting concentration for atotal of 10 dilution points. The serially diluted test antibodies (100μL/well) were added to the cells and incubated for 30 min at 37° C.Cells were washed 2× with FACS staining buffer and AlexaFluor647-conjugated Donkey anti-human secondary antibody (JacksonImmunoresearch) was added and allowed to incubate with the cells for 30min at 4° C. Cells were washed 2× with FACS staining buffer andre-suspended in 100 μL FACS Buffer. Cells were run on BD Celesta usingFACS Diva software and analyzed using FLOWJO. FIG. 16 shows that thePSMA/CD3 bispecific antibodies display differential CD3 cell bindingprofiles detected by flow cytometry.

Binding curves demonstrated in Table 69 below and FIG. 17 were generatedagainst the prostate cell line C4-2B at 37° C. in RPMI media plus 10%fetal bovine serum after 1 hr incubation. Molecule concentrations rangedfrom 500-0 nM over 12-points at 3-fold dilution. Selective binding toPSMA was validated using an isotype control. Values reported in theTable 69 were generated by fitting the data to a four-parameter functionfor ligand binding generating values for y-min, y-max, EC50, and Hill.EC90 were calculated using the equation EC90=(90−(100−90)){circumflexover ( )}(1/Hill)*EC50. All curves exhibited a similar Y-min with anaverage of 7.1+/−1.3E+4 for all curves. See Table 69. None of the Y-minvalues deviated significantly from the average value. The average fittedY-max value was 1.7+/−0.6E+6. Molecule PSMB1069 exhibited a 2-foldhigher binding signal from the average. None of the other moleculesexhibited a significant difference from the average. These moleculesexhibited an average EC50=17+/−12 nM.

TABLE 69 EC50 values of anti-PSMA antibodies binding to PSMA-expressingcell line C4-2B measured by flow cytometry assays. EC50 EC90 MoleculeY-min Y-max (nM) (nM) PSMB1041 7.2E+04 1.1E+06 16 59 PSMB1045 5.3E+041.8E+06 7 248 PSMB1047 7.9E+04 1.6E+06 22 71 PSMB1049 8.4E+04 1.5E+06 540 PSMB1051 6.7E+04 1.5E+06 10 76 PSMB1052 6.8E+04 1.5E+06 13 100PSMB1060 8.6E+04 1.6E+06 11 59 PSMB1068 4.8E+04 1.9E+06 8 247 PSMB10698.7E+04 3.3E+06 44 254 PSMB1075 6.6E+04 1.3E+06 30 156

Binding of anti-PSMA variants/CD3 bispecific on T cells via flowcytometry. C4-2B human prostate tumor cells were washed with DPBS and0.25% trypsin was added to allow cells to detach. Media was added toneutralize trypsin and the cells were transferred to a 15 mL conicalwith DPBS. The cells were centrifuged 1300 rpm for 5 minutes. DPBS wasaspirated and cells were re-suspended in DPBS. The cells were countedusing the Vi-cell XR cell viability analyzer and were plated at100K/well in 100 μL DPBS. The plate was centrifuged 1200 rpm for 3minutes and washed 2× with DPBS. Cells were stained with VioletLive/Dead stain (Thermo-Fisher) and incubated at RT in the dark for 25min. The cells were centrifuged and washed 2× with FACS staining buffer(BD Pharmingen). Test antibodies were diluted to a final startingconcentration of 100 nM in FACS staining buffer and 3-fold serialdilutions were prepared from the starting concentration for a total of10 dilution points. The serially diluted test antibodies (100 μL/well)were added to the cells and incubated for 30 min at 37° C. Cells werewashed 2× with FACS staining buffer and AlexaFluor 647-conjugated Donkeyanti-human secondary antibody (Jackson Immunoresearch) was added andallowed to incubate with the cells for 30 min at 4° C. Cells were washed2× with FACS staining buffer and re-suspended in 100 μL FACS Buffer.Cells were run on BD CELESTA using FACS Diva software and analyzed usingFLOWJO. FIG. 18 shows that PSMA/CD3 bispecific antibodies displaysimilar PSMA cell binding profiles detected by flow cytometry.

Example 10.4: Internalization of PSMA

C4-2B human prostate tumor cells were washed with DPBS and 0.25% trypsinwas added to allow cells to detach. Media was added to neutralizetrypsin and the cells were transferred to a 15 mL conical with DPBS. Thecells were centrifuged 1300 rpm for 5 minutes. DPBS was aspirated andcells were re-suspended in DPBS. The cells were counted using theVi-cell XR cell viability analyzer and were plated at 40K/well in 50 μLPhenol Red-Free PRMI+10% HI FBS. The PSMA/CD3 bispecific or controlantibodies were incubated with IncuCyte® Human Fab-fluor-pH Red Antibodylabeling dye for 15 minutes then 50 μL of conjugatedPSMA/CD3:Fab-fluor-pH Red complex was added to the wells containingC4-2B cells. The plates were placed in an IncuCyte S3® (Essen) at 37° C.with 5% CO2 for 24 hours. The Ab:Fab-fluor complex that is internalizedby the target cells is processed by acidic lysosomes which produces thered fluorogenic signal that is captured and analyzed by the IncuCyte®.FIG. 19 shows that PSMA/CD3 bispecific antibodies internalize but at alesser rate than the transferrin receptor.

Example 10.5: T-Cell Mediated Killing of Bispecific PSMAxCD3 Antibodieson PSMA+ Cells Via Flow Cytometry

Selected bispecific PSMA×CD3 antibodies were assessed for their abilityto mediate T cell mediated killing of prostate cancer cells.

T cell mediated killing of the PSMA×CD3 bispecific antibodies wasmeasured using an assay that indirectly measures cell killing via flowcytometry. Target cell population are identified base on cell viabilityTest samples and controls were prepared at 20 nM in assay medium (10%RPMI, 10% HI FCS). Half log serial dilutions for a 11-point titration ofcompounds in sterile polypropylene plates were prepared. Additionalwells were used for controls without compounds, T cells or tumor cellcontaining wells only in assay medium. C4-2B cells were harvested fromthe cell culture flasks and cells were resuspended in PBS. Cells werestained with 20 pM CFSE for 10 minutes at room temperature. 25 mL of HIFBS was added to stop the staining reaction and the cells werecentrifuged at 300×G for 5 minutes. Cells were diluted to 1×10⁶/ml andthen plated as tumor target cells in 50 μL assay medium for 50,000cells/well into a V-bottom tissue culture treated polystyrene assayplate. 50 μL/well of assay media was added to the control wells that didnot receive tumor cells. Human PAN-T cell vials were thawed in a waterbath set at 37° C. and washed twice by adding 10 ml assay medium andcentrifuging at 400×G for 5 minutes. T cells were resuspended to1×10⁶/mL in assay medium and 50 μL containing 50,000/well were added tothe assay plates containing tumor target cells. 50 μL/well assay mediawas added to the control wells that did not receive tumor cells. 100μL/well of serially diluted antibodies were added to the assay platescontaining cell mixture of target and effector cells. Plates wereincubated at 37° C., 5% CO2 in a humidified cell culture incubator for72 hours.

Following the incubation assay plates were centrifuged at 500×G for 5minutes and medium was removed from the wells. 150 μL DPBS was added toeach well and the plates were centrifuged at 500×G for 5 minutes andmedium was removed from the wells. The cultures were assessed using flowcytometry on the INTELLICYT IQ Plus for viable tumor cells using near IRlive/dead stain. T cell activation was assessed using a brilliantviolet-labeled anti-CD25 MAB. Cells were gated in an FSC vs SSC gate toeliminate debris. Tumor cells were identified as CFSE positive cells. Tcells were identified as CSFE negative cells. Tumor cell viability wascalculated as number of live/dead stain positive tumor cells as apercentage of total CSFE cells. Activated T cells were calculated as thenumber of CD25 positive cells as a percentage of the total number of thelive CFSE negative population. The data for percent dead tumor cells andactivated T cells were plotted vs. antibody concentration in Gene DataScreener using 4 parameter curve fitting to generate EC50 values. Table70 shows EC50 values for T cell activation and tumor cell killing.

TABLE 70 In vitro T cell mediated killing of tumor cells and T cellactivation by bispecific PSMA × CD3 antibodies. Tumor killing T cellactivation Name EC50 [M] EC50 [M] PS3B917 7.31E−12 4.48E−12 PS3B9181.00E−08 1.00E−08 PS3B913 3.11E−10 1.55E−10 PS3B915 1.00E−08 1.00E−08PS3B914 1.00E−08 1.00E−08 PS3B916 1.00E−08 1.00E−08 PS3B919 2.10E−101.87E−10 PS3B921 3.11E−11 2.11E−11 PS3B920 3.20E−09 1.77E−09 PS3B9227.79E−11 5.85E−11 PS3B912 6.51E−11 4.73E−11 PS3B930 4.83E−12 2.40E−12PS3B931 1.00E−08 1.00E−08 PS3B926 4.78E−11 3.45E−11 PS3B928 4.81E−101.84E−10 PS3B927 1.00E−08 1.00E−08 PS3B929 1.00E−08 1.00E−08 PS3B9326.42E−12 1.20E−11 PS3B934 7.24E−12 4.81E−12 PS3B933 4.76E−11 6.01E−11PS3B935 7.16E−12 6.72E−12 PS3B925 7.98E−12 4.59E−12

Example 10.6: T-Cell Mediated Killing of Bispecific PSMAxCD3 Antibodieson PSMA+ Cells Via Incucyte

Select bispecific PSMAxCD3 antibodies were assessed for their ability tomediate T cell mediated killing of prostate cancer cells viaIncuCyte®-based cytotoxic assay.

Healthy donor T cells. PSMA+C4-2B cells stably expressing red nucleardye were generated to be used in the IncuCyte-based cytotoxicity assay.Frozen vials of healthy donor T cells (Biological Specialty Corporation,Colmar, Pa.) were thawed in a 37° C. water bath, transferred to a 15 mLconical tube, and washed once with 5 mL phenol-red-free RPMI/10% HI FBSmedium. The cells were counted using the VIACELL XR cell viabilityanalyzer and the T cells were combined with target cells for a finaleffector T cell to target cell (E: T) ratio of 3:1. The cell mixture wascombined in a 50 mL conical tube. The cell mixture (100 μL/well) wasadded to a clear 96-well flat-bottom plate. Next, the test antibodieswere diluted to a final starting concentration of 60 nM inphenol-red-free RPMI/10% HI FBS medium and 3-fold serial dilutions wereprepared from the starting concentration for a total of 11 dilutionpoints. The serially diluted test antibodies (100 μL/well) were added tothe combined cells. The plates were placed in either an IncuCyte® Zoomor an IncuCyte S3® (Essen) at 37° C. with 5% CO2 for 120 hours. Thetarget cell lines stably express red nuclear dye which is used to trackthe kinetics of target cell lysis. Percent cell growth inhibition(%)=(Initial viable target cell number−Current viable target cellnumber)/Initial viable cell number*100%. Table 71 and FIGS. 20A-20H showcytotoxicity for C4-2B cells with increasing concentrations ofanti-PSMA. Isolated PAN-T cells were co-incubated with PSMA+C4-2B cellsin the presence of bispecific PSMA/T cell redirection antibodies for 120hours.

TABLE 71 Bispecific anti-PSMA/anti-T cell redirection antibodiesevaluated in an IncuCyte ®-based cytotoxicity assay. Cytotoxicity (C4-2Bcells, 3:1 E:T ratio, 5 Day) 30 10 3.3 1.1 0.3 Name nM nM nM nM nMPS3B1352 79% 85% 89% 88% 65% PS3B1356 88% 83% 80% 55% No lysis PS3B135392% 95% 97% 98% 98% PS3B1357 91% 96% 95% 96% 96% PS3B1354 84% 72% 30% NoNo lysis lysis PSMB937 41% No No No No lysis lysis lysis lysis PS3B135594% 95% 96% 97% 92% PS3B1358 88% 94% 93% 90% 68%

Healthy PBMCs. PSMA+C4-2B human prostate tumor cells expressing rednuclear dye were generated to be used in the IncuCyte®-basedcytotoxicity assay. Frozen vials of healthy PBMCs (Hemacare, LosAngeles, Calif.) were thawed in a 37° C. water bath, transferred to a 15mL conical tube, and washed once with 5 mL phenol-red-free RPMI/10% HIFBS medium. The cells were counted using the VIACELL XR cell viabilityanalyzer and the PBMCs were combined with target cells for a final PBMCto target cell (E: T) ratio of 1:1. The cell mixture was combined in a 50 mL conical tube. The cell mixture (100 μL/well) was added to a clear96-well flat-bottom plate. Next, the test antibodies were diluted to afinal starting concentration of 30 nM in phenol-red-free RPMI/10% HI FBSmedium and 3-fold serial dilutions were prepared from the startingconcentration for a total of 11 dilution points. The serially dilutedtest antibodies (100 μL/well) were added to the combined cells. Theplates were placed in either an IncuCyte® Zoom or an IncuCyte S3®(Essen) at 37° C. with 5% CO₂ for 120 hours. The target cell linesstably express red nuclear dye which is used to track the kinetics oftarget cell lysis. Percent cell growth inhibition (%)=(Initial viabletarget cell number−Current viable target cell number)/Initial viablecell number*100%. FIG. 21 shows that the PSMA/CD3 bispecific antibodiesinduce differential C4-2B cytotoxic effects.

Example 10.7: Evaluating Cytokine Induction by Bispecific Anti-PSMAxCD3Antibodies

Select bispecific PSMAxCD3 antibodies were assessed for their ability toinduce cytokine release.

Supernatants collected from the in-vitro cytotoxicity experimentdescribed above were analyzed using the Human Proinflammatory Panel Itissue culture kit (Meso Scale Discovery). Supernatants were thawed onwet ice, spun at 1,500 rpm for 5 minutes at 4° C., then placed on ice.The MULT-SPOT assay plates were pre-washed per the manufacturer'sprotocol. A standard curve was prepared by serial dilution of theprovided calibrators in MSD Diluent 1. The standards and test antibodysamples (254/well) were added to the pre-washed plates. Subsequentincubations and washes were all carried out per manufacturer's protocol.Assay plates were read on the SECTOR Imager 6000. IFNγ concentrationswere quantified for each PSMAxCD3 bispecific antibody evaluated. FIG. 22shows functional cytokine release by T cells activated by PSMAxCD3antibodies.

Example 10.8: T-Cell Mediated Killing of Bispecific PSMAxCD3 Antibodieson PSMA+ Cells Via xCelligence

Select bispecific PSMAxCD3 antibodies were assessed for their ability tomediate T cell mediated killing of prostate cancer cells, C4-2B. C4-2B,a prostate cancer cell line expressing ˜150,000 PSMA/cell was used at a3:1 Effector to Target ratio (E:T), using three PAN-T donors. On day 0of the experiment, xCelligence plates were blanked with 50 μl of growthmedia. Plates were then seeded with 20,000 C4-2B (50 μL out of 0.4×10⁶cells/ml) cells per well. Plates were then incubated on the xCelligencemachine overnight. On day 1 of the experiment, three PAN-T donors wereused to prepare the E:T ratio by adding 50 μL of 1.2×10⁶ cells/mL(60,000 cells). Then 50 μL of the appropriate bispecific antibodies wereadded to the appropriate wells for each plate. CD3×null was used as acontrol. Tumor/target only wells were assigned to be used fornormalization in the percent cytolysis calculation. Final antibodyconcentrations were 50 nM, 10 nM, 2 nM, 0.4 nM, 80 pM and 0 nM. Plateswere then placed in the XCELLIGENCE machine and impedance was recordedevery 15 minutes for 120 hours. Percent cytolysis was calculated on theRTCA software using the equation % cytolysis=[1−(NCI)/(AvgNCIR)]×100,where NCI is the average cell index of the well and AvgNCIR is theaverage cell index of the tumor only reference wells. Table 72summarizes cytolysis for each PSMA×CD3 bispecific molecule over time.

TABLE 72 Summary of % cytolysis at time point 120 hours for all fourbispecific antibodies, for three PAN-T donors, at each doseconcentration. Donor ID Name 50 nM 10 nM 2 nM 0.4 nM 80 pM #1 PS3B1352100% 100% 100% 100%  77% PS3B1353 100% 100% 100% 100% 100% PS3B1356 100%100% 100% 100%  0% PS3B1357 100% 100% 100% 100% 100% #2 PS3B1352 100%100% 100% 100%  0% PS3B1353 100% 100% 100% 100% 100% PS3B1356 100% 100%100%  24%  0% PS3B1357 100% 100% 100% 100% 100% #3 PS3B1352 100% 100%100% 100%  9% PS3B1353 100% 100% 100% 100% 100% PS3B1356 100% 100% 100%100%  0% PS3B1357 100% 100% 100% 100% 100%

It will be appreciated by those skilled in the art that changes could bemade to the embodiments described above without departing from the broadinventive concept thereof. It is understood, therefore, that thisinvention is not limited to the particular embodiments disclosed, but itis intended to cover modifications within the spirit and scope of thepresent invention as defined by the present description.

What is claimed:
 1. An isolated protein comprising an antigen bindingdomain that binds to cluster of differentiation 3ε (CD3ε), wherein theantigen binding domain that binds CD3ε comprises: a. a heavy chaincomplementarity determining region (HCDR) 1, a HCDR2 and a HCDR3 of aheavy chain variable region (VH) of SEQ ID NO: 55 and a light chaincomplementarity determining region (LCDR) 1, a LCDR2 and a LCDR3 of alight chain variable region (VL) of SEQ ID NO: 59; b. the HCDR1, theHCDR2 and the HCDR3 of the VH of SEQ ID NO: 55 and the LCDR1, the LCDR2and the LCDR3 of the VL of SEQ ID NO: 58; c. the HCDR1, the HCDR2 andthe HCDR3 of the VH of SEQ ID NO: 54 and the LCDR1, the LCDR2 and theLCDR3 of the VL of SEQ ID NO: 56; or d. the HCDR1, the HCDR2 and theHCDR3 of the VH of SEQ ID NO: 48 and the LCDR1, the LCDR2 and the LCDR3of the VL of SEQ ID NO: 58; wherein the amino acid in position N106 ofSEQ ID NO: 55, 54, or 48 is optionally substituted with the amino acidselected from the group consisting of A, G, S, F, E, T, R, V, I, Y, L,P, Q, and K, wherein the residue numbering starts from N-terminus of SEQID NO: 55, 54, or
 48. 2. The isolated protein of claim 1, comprising theHCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and the LCDR3 of SEQID NOs: 70, 71, 86, 79, 80, and 81, respectively.
 3. The isolatedprotein of claim 1, comprising the HCDR1, the HCDR2, the HCDR3, theLCDR1, the LCDR2 and the LCDR3 of a. SEQ ID NOs: 70, 71, 72, 79, 80, and81, respectively; b. SEQ ID NOs: 70, 71, 87, 79, 80, and 81,respectively; or c. SEQ ID NOs: 70, 71, 90, 79, 80, and 81,respectively.
 4. The isolated protein of claim 1, wherein the antigenbinding domain that binds CD3ε is a scFv, a (scFv)2, a Fv, a Fab, aF(ab′)2, a Fd, a dAb or a VHH.
 5. The isolated protein of claim 4,wherein the antigen binding domain that binds CD3ε is the Fab.
 6. Theisolated protein of claim 4, wherein the antigen binding domain thatbinds CD3ε is the scFv.
 7. The isolated protein of claim 6, wherein thescFv comprises, from the N- to C-terminus, a VH, a first linker (L1) anda VL (VH-L1-VL) or the VL, the L1 and the VH (VL-L1-VH).
 8. The isolatedprotein of claim 7, wherein the L1 comprises a. about 5-50 amino acids;b. about 5-40 amino acids; c. about 10-30 amino acids; or d. about 10-20amino acids.
 9. The isolated protein of claim 7, wherein the L1comprises an amino acid sequence of SEQ ID NOs: 3-36.
 10. The isolatedprotein of claim 9 wherein the L1 comprises the amino acid sequence ofSEQ ID NO:
 3. 11. The isolated protein of claim 1, wherein the antigenbinding domain that binds CD3ε comprises the VH of SEQ ID NOs: 55, 54,or 48 and the VL of SEQ ID NOs: 59, 58 or
 56. 12. The isolated proteinof claim 11, wherein the antigen binding domain that binds CD3εcomprises: a. the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 59; b.the VH of SEQ ID NO: 55 and the VL of SEQ ID NO: 58; c. the VH of SEQ IDNO: 54 and the VL of SEQ ID NO: 56; d. the VH of SEQ ID NO: 48 and theVL of SEQ ID NO: 58; e. the VH of SEQ ID NO: 88 and the VL of SEQ ID NO:58; or f. the VH of SEQ ID NO: 242 and the VL of SEQ ID NO:
 58. 13. Theisolated protein of claim 1, wherein the antigen binding domain thatbinds CD3ε comprises an amino acid sequence selected from the groupconsisting of SEQ ID NOs: 96, 97, 98, 99, 100, 101, 102, 103, 104, 105,106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119,120, 121, 122, 123, 124, 125, or
 126. 14. The isolated protein of claim1, wherein the isolated protein is a multispecific protein.
 15. Theisolated protein of claim 14, wherein the multispecific protein is abispecific protein.
 16. The isolated protein of claim 14, wherein themultispecific protein is a trispecific protein.
 17. The isolated proteinof claim 1, further comprising an immunoglobulin (Ig) constant region ora fragment of the Ig constant region thereof.
 18. The isolated proteinof claim 17, wherein the fragment of the Ig constant region comprises aFc region.
 19. The isolated protein of claim 17, wherein the fragment ofthe Ig constant region comprises a CH2 domain.
 20. The isolated proteinof claim 17, wherein the fragment of the Ig constant region comprises aCH3 domain.
 21. The isolated protein of claim 17, wherein the fragmentof the Ig constant region comprises a CH2 domain and a CH3 domain. 22.The isolated protein of claim 17, wherein the fragment of the Igconstant region comprises at least portion of a hinge, a CH2 domain anda CH3 domain.
 23. The isolated protein of claim 17, wherein the fragmentof the Ig constant region comprises a hinge, a CH2 domain and a CH3domain.
 24. The isolated protein of claim 17, wherein the antigenbinding domain that binds CD3ε is conjugated to the N-terminus of the Igconstant region or the fragment of the Ig constant region.
 25. Theisolated protein of claim 17, wherein the antigen binding domain thatbinds CD3ε is conjugated to the C-terminus of the Ig constant region orthe fragment of the Ig constant region.
 26. The isolated protein ofclaim 17, wherein the antigen binding domain that binds CD3ε isconjugated to the Ig constant region or the fragment of the Ig constantregion via a second linker (L2).
 27. The isolated protein of claim 26,wherein the L2 comprises the amino acid sequence selected from the groupconsisting of SEQ ID NOs: 3-36.
 28. The isolated protein of claim 14,wherein the multispecific protein comprises an antigen binding domainthat binds an antigen other than CD3ε.
 29. The multispecific antibody ofclaim 14, wherein the antigen is a tumor associated antigen.
 30. Theisolated protein of claim 14, wherein the Ig constant region or thefragment of the Ig constant region is an IgG1, an IgG2, an IgG3 or anIgG4 isotype.
 31. The isolated protein of claim 1, wherein the Igconstant region or the fragment of the Ig constant region comprises atleast one mutation that results in reduced binding of the protein to aFcγ receptor (FcγR).
 32. The isolated protein of claim 31, wherein theat least one mutation that results in reduced binding of the protein tothe FcγR is selected from the group consisting of F234A/L235A,L234A/L235A, L234A/L235A/D265S,V234A/G237A/P238S/H268A/V309L/A330S/P331S, F234A/L235A,S228P/F234A/L235A, N297A, V234A/G237A,K214T/E233P/L234V/L235A/G236-deleted/A327G/P331A/D365E/L358M,H268Q/V309L/A330S/P331S, S267E/L328F, L234F/L235E/D265A,L234A/L235A/G237A/P238S/H268A/A330S/P331S, S228P/F234A/L235A/G237A/P238Sand S228P/F234A/L235A/G236-deleted/G237A/P238S, wherein residuenumbering is according to the EU index.
 33. The isolated protein ofclaim 31, wherein the FcγR is FcγRI, FcγRIIA, FcγRIIB or FcγRIII, or anycombination thereof.
 34. The isolated protein of claim 14, wherein theprotein comprises at least one mutation in a CH3 domain of the Igconstant region.
 35. The isolated protein of claim 34, wherein the atleast one mutation in the CH3 domain of the Ig constant region isselected from the group consisting of T350V, L351Y, F405A, Y407V, T366Y,T366W, T366L, T366L, F405W, T394W, K392L, T394S, T394W, Y407T, Y407A,T366S/L368A/Y407V, L351Y/F405A/Y407V, T366I/K392M/T394W, F405A/Y407V,T366L/K392M/T394W, T366L/K392L/T394W, L351Y/Y407A, L351Y/Y407V,T366A/K409F, T366V/K409F, T366A/K409F, T350V/L351Y/F405A/Y407V andT350V/T366L/K392L/T394W, wherein residue numbering is according to theEU index.
 36. A pharmaceutical composition comprising the isolatedprotein of claim 1 and a pharmaceutically acceptable carrier.
 37. Apolynucleotide encoding the isolated protein of claim
 1. 38. A vectorcomprising the polynucleotide of claim
 37. 39. A host cell comprisingthe vector of claim
 38. 40. A method of producing the isolated proteinof claim 1, comprising culturing the host cell of claim 39 in conditionsthat the protein is expressed, and recovering the protein produced bythe host cell.
 41. An anti-idiotypic antibody binding to the isolatedprotein of claim
 1. 42. An isolated protein of claim 1 comprising anamino acid sequence selected from the group consisting of SEQ ID NOs:127-157.
 43. An isolated protein of any one of claims 1-35 comprising anantibody heavy chain of SEQ ID NO: 224 and antibody light chain of SEQID NO: 226.